Telomerase Targeting in Cancer Treatment: New Developments

Drug Resistance Updates : Reviews and Commentaries in Antimicrobial and Anticancer Chemotherapy. Apr, 1999  |  Pubmed ID: 11504478

Telomerase, a ribonucleoprotein expressed in 85% of advanced cancers but not in most somatic cells, compensates for telomeric DNA erosion and as such stabilizes cell immortality. Telomerase inhibition might restore mortality in tumor cells. Recent progress is illustrated in studies on telomerase and telomere targeting with differentiation induction, reverse transcriptase inhibitors, promoter down regulation, antisense inhibition, and blockage of telomere/telomerase interactions. Also, new developments are described indicating that anti-telomerase treatment can induce apoptosis in tumor cells and can chemosensitize drug-resistant cell lines. Implications of these findings for anti-telomerase-based therapeutic applications, in particular in combination therapies, are discussed. Copyright 1999 Harcourt Publishers Ltd.

Molecular and Clinical Characteristics of MSH6 Variants: an Analysis of 25 Index Carriers of a Germline Variant

American Journal of Human Genetics. Jan, 2002  |  Pubmed ID: 11709755

The MSH6 gene is one of the mismatch-repair genes involved in hereditary nonpolyposis colorectal cancer (HNPCC). Three hundred sixteen individuals who were known or suspected to have HNPCC were analyzed for MSH6 germline mutations. For 25 index patients and 8 relatives with MSH6 variants, molecular and clinical features are described. For analysis of microsatellite instability (MSI), the five consensus markers were used. Immunohistochemical analysis of the MLH1, MSH2, and MSH6 proteins was performed. Five truncating MSH6 mutations, of which one was detected seven times, were found in 12 index patients, and 10 MSH6 variants with unknown pathogenicity were found in 13 index patients. Fourteen (54%) of 26 colorectal cancers (CRCs) and endometrial cancers showed no, or only weak, MSI. Twelve of 18 tumors of truncating-mutation carriers and 3 of 17 tumors of missense-mutation carriers showed loss of MSH6 staining. Six of the families that we studied fulfilled the original Amsterdam criteria; most families with MSH6, however, were only suspected to have HNPCC. In families that did not fulfill the revised Amsterdam criteria, the prevalence of MSH6 variants is about the same as the prevalence of those in MLH1/MSH2. Endometrial cancer and/or atypical hyperplasia were diagnosed in 8 of 12 female carriers of MSH6 truncating mutations. Most CRCs were localized distally in the colon. Although, molecularly, missense variants are labeled as doubtfully pathogenic, clinical data disclose a great resemblance between missense-variant carriers and truncating-mutation carriers. We conclude that, in all patients suspected to have HNPCC, MSH6-mutation analysis should be considered. Neither MSI nor immunohistochemistry should be a definitive selection criterion for MSH6-mutation analysis.

Vulvar Melanoma: is There a Role for Sentinel Lymph Node Biopsy?

Cancer. Jan, 2002  |  Pubmed ID: 11905414

The objective of this study was to evaluate the author's recent, preliminary experience with the sentinel lymph node procedure in patients with vulvar melanoma and to compare this experience with treatment and follow-up of patients with vulvar melanomas who were treated previously at their institution.

Vulvar Carcinoma. The Price of Less Radical Surgery

Cancer. Dec, 2002  |  Pubmed ID: 12436439

The objective of this study was to determine whether modifications in the treatment of patients with vulvar carcinoma influence the rates of recurrence and survival.

Sentinel Node Techniques in Cancer of the Vulva

Current Women's Health Reports. Feb, 2003  |  Pubmed ID: 12521546

The sentinel lymph node procedure, with the combined technique (preoperative lymphoscintigraphy with (99m)Technetium-labeled Nanocolloid and Patente Blue V ), is a promising staging technique for patients with vulvar cancer. The clinical implementation of the sentinel lymph node procedure and the role of additional histopathologic techniques of the sentinel lymph nodes are under investigation.

The Serum Concentration of Human Kallikrein 10 Represents a Novel Biomarker for Ovarian Cancer Diagnosis and Prognosis

Cancer Research. Feb, 2003  |  Pubmed ID: 12591730

Human kallikrein 10 (hK10) is a secreted serine protease that is highly expressed in ovarian tissue. We hypothesized that hK10 might represent a novel serological marker for ovarian cancer. We quantified by immunoassay, hK10 in sera from 97 normal women (controls), 141 patients with benign gynecologic diseases, and 146 patients with ovarian cancer. We then examined the diagnostic and prognostic value of this measurement in ovarian cancer. We found that normal serum hK10 ranged from 50 to 1040 ng/liter (mean = 439 ng/liter). hK10 concentration is significantly elevated in serum of presurgical ovarian cancer patients (range: 106-11,746 ng/liter; mean = 1067 ng/liter) but not in serum of patients with benign gynecologic diseases (range: 120-1200 ng/liter; mean = 447 ng/liter). When a cutoff of 700 ng/liter was selected (diagnostic specificity = 90%), the diagnostic sensitivity for ovarian cancer is 54%. About 35% of CA125-negative ovarian cancer patients (CA125 < 23 kU/liter) were hK10 positive at 90% specificity. In patients with stage I/II ovarian cancer, use of these two markers in combination results in a 21% increase in sensitivity, at 90% specificity, compared with CA125 alone. High serum hK10 was strongly associated with serous epithelial type, late-stage, advanced grade, large residual tumor (>1 cm), suboptimal debulking, and no response to chemotherapy (all Ps < 0.001). In univariate Cox survival analysis, high serum hK10 is associated with increased risk for relapse and death (hazard ratio = 2.59 and 3.15, respectively, P

Human Kallikrein 6 (hK6): a New Potential Serum Biomarker for Diagnosis and Prognosis of Ovarian Carcinoma

Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Mar, 2003  |  Pubmed ID: 12637468

The discovery of new ovarian cancer biomarkers that are suitable for early disease diagnosis and prognosis may ultimately lead to improved patient management and outcomes.

HLA Class I Alleles and Cervical Neoplasia

The Journal of Infectious Diseases. May, 2003  |  Pubmed ID: 12721951

Re: Further Data on the Usefulness of Sentinel Lymph Node Identification and Ultrastaging in Vulvar Squamous Cell Carcinoma. Gynecol Oncol. 2003 Jan;88(1):29-34

Gynecologic Oncology. Sep, 2003  |  Pubmed ID: 14515864

Gynecologic Screening in Hereditary Nonpolyposis Colorectal Cancer

Gynecologic Oncology. Oct, 2003  |  Pubmed ID: 14529665

In hereditary nonpolyposis colorectal cancer (HNPCC), women with a mismatch repair (MMR) gene mutation have a cumulative lifetime risk of 25-50% for endometrial cancer and 8-12% for ovarian cancer. Therefore, female members of HNPCC families are offered an annual gynecologic and transvaginal ultrasound (TVU) examination and serum level CA 125 analysis. The aim of the present study was to evaluate our 10-year experience with this screening program.

Telomerase in Relation to Expression of P53, C-Myc and Estrogen Receptor in Ovarian Tumours

International Journal of Oncology. Nov, 2003  |  Pubmed ID: 14532990

Telomerase activity and its subunits (hTERC, hTERT mRNA) were evaluated in ovarian tumours in relation to the expression of p53, c-Myc and estrogen receptor (ER). Furthermore, relations between telomerase activity, hTERC and hTERT with known clinicopathologic prognostic factors and survival in patients with malignant tumours was investigated. Telomerase activity was determined with TRAP, hTERC and hTERT with RT-PCR, while p53, c-Myc and ER expression with immunohistochemistry. Telomerase activity and hTERT mRNA were more frequently observed in malignant ovarian tumours compared to borderline and benign tumours, whereas hTERC was present in all tumour types. p53 and c-Myc were more frequently detected in malignant compared to borderline and benign tumours. Telomerase activity was positively related to hTERT mRNA, p53 and c-Myc expression, but not to hTERC and ER expression. In malignant tumours, hTERC levels were related to tumour stage, while telomerase activity and hTERT mRNA expression were not related to any clinicopathologic feature. Tumour stage, differentiation grade, residual tumour after first laparotomy and presence of ascites were related to (progression free) survival, whereas telomerase activity or its subunits were not. In conclusion, these data suggest that p53 expression (e.g. p53 mutation) as well as c-Myc expression may have a role in regulation of telomerase activity in ovarian tumours.

Toward New Strategies to Select Young Endometrial Cancer Patients for Mismatch Repair Gene Mutation Analysis

Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Dec, 2003  |  Pubmed ID: 14645426

To determine the frequency of mismatch repair (MMR) gene germline mutations in endometrial cancer patients who were diagnosed at less than 50 years of age; to relate the presence of mutations to family history, histopathologic data, presence of tumor microsatellite instability (MSI), and immunostaining; and to formulate criteria for genetic testing in these patients.

The Value of Routine Follow-up in Patients Treated for Carcinoma of the Vulva

Cancer. Dec, 2003  |  Pubmed ID: 14669281

Vulvar carcinoma patients traditionally are offered follow-up after their primary treatment because earlier diagnosis of recurrent disease is believed to improve chances for curative treatment. The objective of the current study was to determine the value of a strict routine follow-up protocol for the detection of recurrences in a large series of patients who were treated for carcinoma of the vulva.

Elevated HTERT MRNA Levels: A Potential Determinant of Bronchial Squamous Cell Carcinoma (in Situ)

International Journal of Cancer. Journal International Du Cancer. Apr, 2004  |  Pubmed ID: 14961580

Expression levels of hTERT mRNA were investigated by RT-PCR in tissue specimens of patients with (Group A) and without (Group B) clinically overt bronchial carcinoma, respectively. Bronchial carcinoma (n = 9) and distant normal (n = 9) specimens were analyzed in Group A. The chance of carcinoma seemed to increase with increasing hTERT mRNA levels (OR = 6.04, 95% CI = 1.02-37). Group B was comprised of 21 patients who underwent autofluorescence bronchoscopy. After analysis of 66 bronchial biopsies the chance of prevalent carcinoma in situ or carcinoma increased with increasing hTERT mRNA levels (OR = 6.19, 95% CI = 1.55-25). Variables like age, gender, smoking history, history of cancer within the airways or the degree of lymphocyte infiltrate in the specimens did not modify this relation. In 7 Group B patients in whom bronchial cancer was diagnosed during follow-up, biopsies taken before cancer diagnosis from both the area of the newly developed tumor and distantly from this area had been analyzed for hTERT expression. The median hTERT mRNA level in the biopsies from the area of future cancer was significantly higher than in biopsies taken from distant sites (p < 0.03). These data indicate that elevated hTERT mRNA is associated with an increased relative risk of prevalent and incident bronchial squamous cell carcinoma (in situ).

Letter Commenting on "Patholologic Finding in Prophylactic Oophorectomy Specimens in High-risk Women" (87:52-6) by Pamela Paley, Et Al. (, Fax: +31-206-685-9607)

Gynecologic Oncology. May, 2004  |  Pubmed ID: 15099985

Modern Management of Vulvar Cancer

Current Opinion in Obstetrics & Gynecology. Feb, 2004  |  Pubmed ID: 15128010

The radical surgical approach in the treatment of vulvar cancer patients has led to a favourable prognosis for the majority of patients with early stage vulvar cancer. However, morbidity is impressive, leading to more individualized treatment. The authors have reviewed the most recent literature on the pros and cons of the modifications in treatment, including surgery and primary radiotherapy, for primary squamous cell carcinoma of the vulva and vulvar melanoma.

Detecting Cervical Cancer by Quantitative Promoter Hypermethylation Assay on Cervical Scrapings: a Feasibility Study

Molecular Cancer Research : MCR. May, 2004  |  Pubmed ID: 15192122

Current morphology-based cervical cancer screening is associated with significant false-positive and false-negative results. Tumor suppressor gene hypermethylation is frequently present in cervical cancer. It is unknown whether a cervical scraping reflects the methylation status of the underlying epithelium, and it is therefore unclear whether quantitative hypermethylation specific PCR (QMSP) on cervical scrapings could be used as a future screening method augmenting the current approach. Cervical scrapings and paired fresh frozen cervical tissue samples were obtained from 53 cervical cancer patients and 45 controls. All scrapings were morphologically scored and analyzed with QMSP for the genes APC, DAPK, MGMT, and GSTP1. To adjust for DNA input, hypermethylation ratios were calculated against DNA levels of a reference gene. Hypermethylation ratios of paired fresh frozen tissue samples and scrapings of cervical cancer patients and controls were strongly related (Spearman correlation coefficient, 0.80 for APC, 0.98 for DAPK, and 0.83 for MGMT; P < 0.001). More cervical cancer patients than controls were DAPK positive (P < 0.001). When cutoff levels for ratios were defined to be above the highest ratio observed in controls, QMSP in cervical scrapings identified 32 (67%) of 48 cervical cancer patients. This feasibility study demonstrates that QMSP on cervical scrapings holds promise as a new diagnostic tool for cervical cancer. The addition of more genes specifically methylated in cervical cancer will further improve the assay.

Three Biomarkers Identified from Serum Proteomic Analysis for the Detection of Early Stage Ovarian Cancer

Cancer Research. Aug, 2004  |  Pubmed ID: 15313933

Early detection remains the most promising approach to improve long-term survival of patients with ovarian cancer. In a five-center case-control study, serum proteomic expressions were analyzed on 153 patients with invasive epithelial ovarian cancer, 42 with other ovarian cancers, 166 with benign pelvic masses, and 142 healthy women. Data from patients with early stage ovarian cancer and healthy women at two centers were analyzed independently and the results cross-validated to discover potential biomarkers. The results were validated using the samples from two of the remaining centers. After protein identification, biomarkers for which an immunoassay was available were tested on samples from the fifth center, which included 41 healthy women, 41 patients with ovarian cancer, and 20 each with breast, colon, and prostate cancers. Three biomarkers were identified as follows: (a) apolipoprotein A1 (down-regulated in cancer); (b) a truncated form of transthyretin (down-regulated); and (c) a cleavage fragment of inter-alpha-trypsin inhibitor heavy chain H4 (up-regulated). In independent validation to detect early stage invasive epithelial ovarian cancer from healthy controls, the sensitivity of a multivariate model combining the three biomarkers and CA125 [74% (95% CI, 52-90%)] was higher than that of CA125 alone [65% (95% CI, 43-84%)] at a matched specificity of 97% (95% CI, 89-100%). When compared at a fixed sensitivity of 83% (95% CI, 61-95%), the specificity of the model [94% (95% CI, 85-98%)] was significantly better than that of CA125 alone [52% (95% CI, 39-65%)]. These biomarkers demonstrated the potential to improve the detection of early stage ovarian cancer.

Preoperative Sensitivity and Specificity for Early-stage Ovarian Cancer when Combining Cancer Antigen CA-125II, CA 15-3, CA 72-4, and Macrophage Colony-stimulating Factor Using Mixtures of Multivariate Normal Distributions

Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Oct, 2004  |  Pubmed ID: 15381683

In CA-125-based ovarian cancer screening trials, overall specificity and screening sensitivity of ultrasound after an elevated CA-125 exceeded 99.6% and 70%, respectively, thereby yielding a positive predictive value (PPV) exceeding 10%. However, sensitivity for early-stage disease was only 40%. This study aims to increase preoperative sensitivity for early-stage ovarian cancer while maintaining the annual referral rate to ultrasound at 2% by combining information across CA-125II, CA 15-3, CA 72-4, and macrophage colony-stimulating factor (M-CSF). For direct comparisons between marker panels, all sensitivity results correspond to a 98% fixed first-line specificity (referral rate 2%).

Methylenetetrahydrofolate Reductase (MTHFR) and Susceptibility for (pre)neoplastic Cervical Disease

Human Genetics. Mar, 2005  |  Pubmed ID: 15635481

Methylenetetrahydrofolate reductase (MTHFR) is a critical enzyme regulating the metabolism of folate and methionine. The potential influence of MTHFR activity on DNA methylation and on the availability of uridylates and thymidylates for DNA synthesis and repair presents MTHFR as a candidate for being a cancer-predisposing gene. In the present study, we have examined a large study population to determine whether the C677T polymorphism at the MTHFR locus affects susceptibility for cervical cancer or its precursor, cervical intraepithelial neoplasia (CIN). In addition, we have investigated whether this polymorphism is causal, and not merely associated, by typing microsatellite markers in the region surrounding the MTHFR gene. A total of 311 CIN and 695 cervical cancer patients and 115 family-based and 586 unrelated controls was analysed. Association analysis showed a decreased cervical cancer risk for individuals heterozygous or homozygous for the T-allele, both for squamous cell carcinoma (heterozygous odds ration [OR] 0.66 [0.51-0.86]; homozygous OR 0.76 [0.49-1.16]) and adenocarcinoma (heterozygous OR 0.71 [0.49-1.03]; homozygous OR 0.34 [0.14-0.81]). No difference was found for high grade CIN (heterozygous OR 1.03 [0.76-1.40]; homozygous OR 0.91 [0.54-1.55]). A microsatellite haplotype containing the C allele was associated with an increased risk for cervical cancer and CIN (both among squamous cell carcinomas, adenocarcinomas and CIN II-III; OR = 2.61 [1.59-4.27]). Our study thus lends further support to the hypothesis that the MTHFR C677T polymorphism is involved in susceptibility cervical cancer but also illustrates that, despite the large sample size analysed, still larger studies are needed to establish fully the nature of this association.

Fas and Fas Ligand in Cyst Fluids, Serum and Tumors of Patients with Benign and (borderline) Malignant Ovarian Tumors

International Journal of Oncology. Feb, 2005  |  Pubmed ID: 15645122

Drug resistance in ovarian cancer treatment urges the exploration of new targets for drugs against this malignancy. Fas is a cell membrane receptor which, after engagement with Fas ligand (FasL), triggers apoptotic death. In this study Fas and FasL levels in cyst fluids and sera of patients with benign, borderline and malignant ovarian tumors and in corresponding tumors are determined. Fas and FasL were determinded by ELISA and immunohistochemistry in 30 patients with benign, 5 patients with borderline and 24 patients with malignant epithelial ovarian tumors. In serum there were no differences in median Fas levels, while median FasL levels were higher in healthy women (p=0.02). In malignant cyst fluids, median Fas levels where higher compared to benign cyst fluids (p<0.01). FasL immunostaining was more frequent in malignant ovarian tumors (p=0.002). In conclusion, serum Fas or FasL levels do not seem useful markers. Elevated Fas and equal FasL levels in malignant cyst fluids, suggest an increased production of Fas, and not of FasL by malignant cells. High expression of both Fas and FasL, in malignant ovarian tumors present Fas/FasL as an interesting route to explore for innovative cancer therapy.

HLA Genes and Other Candidate Genes Involved in Susceptibility for (pre)neoplastic Cervical Disease

International Journal of Oncology. Mar, 2005  |  Pubmed ID: 15703836

This review focuses on common and genetic risk factors such as HLA and other genes that may be involved in susceptibility for (pre)neoplastic cervical disease. The goal of this review is the evaluation of polymorphisms that are either associated with cervical intraepithelial neoplasia (CIN) and/or cervical cancer. A pooled analysis was performed for DQA1, DQB1 and DRB1 alleles and 10 other genes that have been evaluated in more than one study. An association, either an increased or a decreased risk, with CIN and cervical cancer at a 5% significance level was found for 15 HLA II alleles. Four polymorphisms (Tp53, IL-10, CYP2D6 and the MTHFR) exhibited an increased CIN and cervical cancer risk. However, only the pooled analysis of the DQB1 alleles, the HLA-DR specificities and Tp53 genes had sufficiently large sample sizes to confirm or exclude the proposed association. Our data indicate that further analysis in larger sample sizes, especially for genes other than the HLA genes, is necessary to describe the exact relations between these genes and susceptibility for CIN and cervical cancer with an adequate power.

Lymphvascular Space Involvement: an Independent Prognostic Factor in Endometrial Cancer

Gynecologic Oncology. Mar, 2005  |  Pubmed ID: 15721428

To evaluate whether lymphvascular space involvement (LVSI) is a risk factor for relapse of disease and lymph node metastasis in endometrial cancer.

Preoperative Serum Squamous Cell Carcinoma Antigen Levels in Clinical Decision Making for Patients with Early-stage Cervical Cancer

Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Mar, 2005  |  Pubmed ID: 15735121

To prevent morbidity associated with double modality treatment, early-stage cervical cancer patients should only be offered surgery when there is a low likelihood for adjuvant radiotherapy. We analyzed whether serum squamous cell carcinoma antigen (SCC-ag) analysis allows better preoperative identification of patients with a low likelihood for adjuvant radiotherapy than currently used clinical parameters.

Sensitivity to Fas-mediated Apoptosis in High-risk HPV-positive Human Cervical Cancer Cells: Relationship with Fas, Caspase-8, and Bid

Gynecologic Oncology. May, 2005  |  Pubmed ID: 15863130

Binding of Fas ligand or agonistic anti-Fas antibody to the death receptor Fas can activate a caspase-cascade resulting in apoptosis. In the present study, the functionality of the Fas pathway was studied in human cervical cancer cells with different HPV and p53 status.

Clinical Potential of Inhibitors of Survival Pathways and Activators of Apoptotic Pathways in Treatment of Cervical Cancer: Changing the Apoptotic Balance

The Lancet Oncology. Aug, 2005  |  Pubmed ID: 16054570

Cervical cancer is the most common gynaecological malignant disorder worldwide. The best possible treatment of locally advanced cervical cancer is a combination of radiation and cisplatin-based chemotherapy. However, 5-year overall survival is still only 52%. To improve treatment results, research should focus on the discovery of innovative drug strategies. Drugs directed at inducing tumour-cell apoptosis are regarded as important treatment modalities. Here, we present an overview of the molecular options that can change the apoptotic balance in cervical cancer, through increasing death-receptor-mediated apoptosis, the use of proteasome inhibitors, short interfering RNAs, or non-steroidal anti-inflammatory drugs (NSAIDs). Furthermore, the potential of attacking prosurvival signalling through the epidermal-growth-factor receptor and insulin-like-growth-factor receptor to support the apoptotic process is discussed. Additional research is needed to elucidate the clinical potential of these compounds in the treatment of cervical cancer.

Distinct Regulation and Impact of Type 1 T-cell Immunity Against HPV16 L1, E2 and E6 Antigens During HPV16-induced Cervical Infection and Neoplasia

International Journal of Cancer. Journal International Du Cancer. Feb, 2006  |  Pubmed ID: 16108057

Cervical cancer is the possible outcome of a genital infection with high-risk human papillomavirus type 16 (HPV16) and is preceded by a phase of persistent HPV infection during which the host immune system fails to eliminate the virus. Our previous work showed that failure is reflected by the absence of type 1 T-cell immunity against HPV16 early antigens E2 and E6 in patients with HPV16+ cervical lesions. We now show that a majority of both patients with cervical lesions and healthy subjects display HPV16 L1 peptide-specific type 1 T-cell responses with similar magnitude. The T-cell response in patients was directed at a broad range of peptides within L1, suggesting that during persistent or repeated exposure to HPV16 L1, the immune system maximizes its efforts to counter the viral challenge. Unlike the type 1 T-cell responses against HPV16 early antigens E2 and E6, type 1 T-cell immunity against L1 does not correlate with health or disease. This argues that T-cell responses against early and late HPV16 antigens essentially differ in the manner in which they are induced and regulated, as well as in their impact on the subsequent stages of HPV16-induced cervical disease.

Proteasome Inhibitor MG132 Sensitizes HPV-positive Human Cervical Cancer Cells to RhTRAIL-induced Apoptosis

International Journal of Cancer. Journal International Du Cancer. Apr, 2006  |  Pubmed ID: 16287099

In cervical carcinogenesis, the p53 tumor suppressor pathway is disrupted by HPV (human papilloma virus) E6 oncogene expression. E6 targets p53 for rapid proteasome-mediated degradation. We therefore investigated whether proteasome inhibition by MG132 could restore wild-type p53 levels and sensitize HPV-positive cervical cancer cell lines to apoptotic stimuli such as rhTRAIL (recombinant human TNF-related apoptosis inducing ligand). In a panel of cervical cancer cell lines, CaSki was highly, HeLa intermediate and SiHa not sensitive to rhTRAIL-induced apoptosis. MG132 strongly sensitized HeLa and SiHa to rhTRAIL-induced apoptosis in a caspase-dependent and time-dependent manner. MG132 massively induced TRAIL receptor DR4 and DR5 membrane expression in HeLa, whereas in SiHa only DR5 membrane expression was upregulated from almost undetectable to high levels. Antagonistic DR4 antibody partially inhibited apoptosis induction by rhTRAIL and MG132 in HeLa but had no effect on apoptosis in SiHa. Inhibition of E6-mediated p53 proteasomal degradation by MG132 resulted in elevated levels of active p53 as demonstrated by p53 small interfering RNA (siRNA) sensitive p21 upregulation. Although p53 siRNA partially inhibited MG132-induced DR5 upregulation in HeLa and SiHa, no effect on rhTRAIL-induced apoptosis was observed. MG132 plus rhTRAIL enhanced caspase 8 and caspase 3 activation and concomitant cleavage of X-linked inhibitor of apoptosis (XIAP), particularly in HeLa. In addition, caspase 9 activation was only observed in HeLa. Downregulation of XIAP using siRNA in combination with rhTRAIL induced high levels of apoptosis in HeLa, whereas MG132 had to be added to the combination of XIAP siRNA plus rhTRAIL to induce apoptosis in SiHa. In conclusion, proteasome inhibition sensitized HPV-positive cervical cancer cell lines to rhTRAIL independent of p53. Our results indicate that not only DR4 and DR5 upregulation but also XIAP inactivation contribute to rhTRAIL sensitization by MG132 in cervical cancer cell lines. Combining proteasome inhibitors with rhTRAIL may be therapeutically useful in cervical cancer treatment.

Surgery by Consultant Gynecologic Oncologists Improves Survival in Patients with Ovarian Carcinoma

Cancer. Feb, 2006  |  Pubmed ID: 16369985

Consultant gynecologic oncologists from the regional Comprehensive Cancer Center assisted community gynecologists in the surgical treatment of patients with ovarian carcinoma when they were invited. For this report, the authors evaluated the effects of primary surgery by a gynecologic oncologist on treatment outcome.

Patterns and Frequency of Recurrences of Squamous Cell Carcinoma of the Vulva

Gynecologic Oncology. Oct, 2006  |  Pubmed ID: 16631238

To analyze patterns and frequency of recurrences of squamous cell carcinoma (SCC) of the vulva after wide local excision (WLE) and superficial inguinal lymphadenectomy with separate incisions and to identify prognostic factors for the development of recurrences.

Assessment of Gene Promoter Hypermethylation for Detection of Cervical Neoplasia

International Journal of Cancer. Journal International Du Cancer. Oct, 2006  |  Pubmed ID: 16736496

Current cervical cancer screening is based on morphological assessment of Pap smears and associated with significant false negative and false positive results. Previously, we have shown that detection of hypermethylated genes in cervical scrapings using quantitative methylation-specific PCR (QMSP) is a promising tool for identification of squamous cell cervical cancer. Aim of the present pilot-study was to evaluate presence of hypermethylated genes in cervical carcinogenesis, both in squamous cell as well as adenocarcinomas. Cervical scrapings were obtained from 30 patients diagnosed with cervical cancer (20 squamous cell carcinomas and 10 adenocarcinomas) and 19 women with histologically normal cervices. The scraped cells were used for determination of promoter hypermethylation by QMSP for 12 genes and for morphological assessment. Overall, CALCA, DAPK, ESR1, TIMP3, APC and RAR-beta2 promoters were significantly more often hypermethylated in cancers than in controls, while adenocarcinomas were more often hypermethylated above the highest control ratio for APC, TIMP3 and RASSF1A promoters. Combining 4 genes (CALCA, DAPK, ESR1 and APC) yielded a sensitivity of 89% (with all adenocarcinomas identified), equal to cytomorphology (89%) and high-risk human papilloma virus (Hr-HPV; 90%). The 4-gene QMSP proved theoretically superior to cytomorphology as well as Hr-HPV in specificity (100% vs. 83 and 68%, respectively), because cytology identified 3 controls as moderate or severe dyskaryosis and 6 controls were positive for Hr-HPV. In conclusions, QMSP of 4 gene promoters combined appears to have comparable sensitivity and potentially better specificity in comparison to "classic" cytomorphological assessment and Hr-HPV detection. QMSP holds promise as a new diagnostic tool for both squamous cell carcinoma and adenocarcinoma of the cervix.

Gynecologic Examination and Cervical Biopsies After (chemo) Radiation for Cervical Cancer to Identify Patients Eligible for Salvage Surgery

International Journal of Radiation Oncology, Biology, Physics. Nov, 2006  |  Pubmed ID: 16904839

The aim of this study was to evaluate efficacy of gynecologic examination under general anesthesia with cervical biopsies after (chemo) radiation for cervical cancer to identify patients with residual disease who may benefit from salvage surgery.

A Biological Question and a Balanced (orthogonal) Design: the Ingredients to Efficiently Analyze Two-color Microarrays with Confirmatory Factor Analysis

BMC Genomics. 2006  |  Pubmed ID: 16968534

Factor analysis (FA) has been widely applied in microarray studies as a data-reduction-tool without any a-priori assumption regarding associations between observed data and latent structure (Exploratory Factor Analysis).A disadvantage is that the representation of data in a reduced set of dimensions can be difficult to interpret, as biological contrasts do not necessarily coincide with single dimensions. However, FA can also be applied as an instrument to confirm what is expected on the basis of pre-established hypotheses (Confirmatory Factor Analysis, CFA). We show that with a hypothesis incorporated in a balanced (orthogonal) design, including 'SelfSelf' hybridizations, dye swaps and independent replications, FA can be used to identify the latent factors underlying the correlation structure among the observed two-color microarray data. An orthogonal design will reflect the principal components associated with each experimental factor. We applied CFA to a microarray study performed to investigate cisplatin resistance in four ovarian cancer cell lines, which only differ in their degree of cisplatin resistance.

An Overview of Innovative Techniques to Improve Cervical Cancer Screening

Cellular Oncology : the Official Journal of the International Society for Cellular Oncology. 2006  |  Pubmed ID: 17167177

Although current cytomorphology-based cervical cancer screening has reduced the incidence of cervical cancer, Pap-smears are associated with high false positive and false negative rates. This has spurred the search for new technologies to improve current screening. New methodologies are automation of Pap-smear analysis, addition of new biological or molecular markers to traditional cytology or using these new markers to replace the current screening method. In this overview we will summarize data on cervical cancer epidemiology and etiology and the current cervical cancer screening approach. Available data on new screening approaches, such as quantitative cytochemistry, detection of loss of heterozygosity (LOH) and hypermethylation analysis will be reviewed. We discuss the potential of these approaches to replace or augment current screening. When available, data on cost-effectiveness of certain approaches will be provided. In short, Human Papillomavirus (HPV) DNA detection stands closest to implementation in nation-wide screening programs of all markers reviewed. However, specificity is low in women aged <35 years and the psychological effects of knowledge of HPV positivity in absence of cervical (pre) malignant disease are important drawbacks. In our opinion the results of large clinical trials should be awaited before proceeding to implement HPV DNA detection. New technologies based on molecular changes associated with cervical carcinogenesis might result in comparable sensitivity, but improved specificity. Hypermethylation analysis is likely to be more objective to identify patients with high grade squamous intra-epithelial lesions (HSIL) or invasive cancer with a higher specificity than current cytomorphology based screening.

Sulindac Treatment in Hereditary Non-polyposis Colorectal Cancer

European Journal of Cancer (Oxford, England : 1990). May, 2007  |  Pubmed ID: 17434727

Non-steroidal anti-inflammatory drugs, e.g. sulindac have been extensively studied for chemoprevention in familial adenomatous polyposis, but not in hereditary non-polyposis colorectal cancer (HNPCC). We evaluated these effects in HNPCC using surrogate end-points for cancer risk. In a randomised double-blind cross-over study, 22 subjects (9 female; age 30-66 years, mean 44), all ascertained or probable mutation carriers for HNPCC, were included. Sulindac 150 mg b.i.d. and placebo were given for 4 weeks each, with 4 weeks in between, with biopsies taken from ascending, transverse and sigmoid colon and rectum by colonoscopy after both periods. Proliferation was determined by Ki-67 staining and apoptosis by staining of cytokeratin 18 cleavage products. Expression of cyclins B1, D3 and E and p21, p27, bax, bcl2 and cox-2 was studied immunohistochemically. Proliferation was higher during sulindac treatment than drug placebo treatment in ascending and transverse colon, but not in sigmoid and rectum. Apoptosis was not affected. Besides an increase in cyclin D3, no differences were found in expression of regulating proteins in the proximal colon. CONCLUSION: Sulindac induces an increase in epithelial cell proliferation in the proximal colon of subjects with HNPCC. Since colorectal cancer predominantly arises in the proximal colon in HNPCC, these results cast doubts on the potential chemopreventive effects of sulindac in HNPCC.

Serum Cytokine Profiling As a Diagnostic and Prognostic Tool in Ovarian Cancer: a Potential Role for Interleukin 7

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Apr, 2007  |  Pubmed ID: 17438097

To evaluate if serum cytokine levels could be used as diagnostic or prognostic markers in ovarian cancer.

Beware of Amenorrhea During Tamoxifen: It May Be a Wolf in Sheep's Clothing

Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Aug, 2007  |  Pubmed ID: 17704431

Profiling Studies in Ovarian Cancer: a Review

The Oncologist. Aug, 2007  |  Pubmed ID: 17766655

Ovarian cancer is a heterogeneous disease with respect to histopathology, molecular biology, and clinical outcome. In advanced stages, surgery and chemotherapy result in an approximately 25% overall 5-year survival rate, pointing to a strong need to identify subgroups of patients that may benefit from targeted innovative molecular therapy. This review summarizes: (a) microarray research identifying gene-expression profiles in ovarian cancer; (b) the methodological flaws in the available microarray studies; and (c) applications of pathway analysis to define new molecular subgroups. Microarray technology now permits the analysis of expression levels of thousands of genes. So far seven studies have aimed to identify a genetic profile that can predict survival/clinical outcome and/or response to platinum-based therapy. To date, the clinical evidence of prognostic microarray studies has only reached the level of small retrospective studies, and there are other issues that may explain the nonreproducibility among the reported prognostic profiles, such as overfitting, technical platform differences, and accuracy of measurements. We consider pathway analysis a promising new strategy. The accumulation of small differential expressions within a meaningful molecular regulatory network might lead to a critical threshold level, resulting in ovarian cancer. Microarray technologies have already provided valuable expression data for classifying ovarian cancer and the first clues about which molecular changes in ovarian cancer could be exploited in new treatment strategies. Further improvements in technology as well as in study design, combined with pathway analysis, will allow us to detect even more subtle tumor expression differences among subgroups of ovarian cancer patients. Disclosure of potential conflicts of interest is found at the end of this article.

Evidence Based Selection of Housekeeping Genes

PloS One. 2007  |  Pubmed ID: 17878933

For accurate and reliable gene expression analysis, normalization of gene expression data against housekeeping genes (reference or internal control genes) is required. It is known that commonly used housekeeping genes (e.g. ACTB, GAPDH, HPRT1, and B2M) vary considerably under different experimental conditions and therefore their use for normalization is limited. We performed a meta-analysis of 13,629 human gene array samples in order to identify the most stable expressed genes. Here we show novel candidate housekeeping genes (e.g. RPS13, RPL27, RPS20 and OAZ1) with enhanced stability among a multitude of different cell types and varying experimental conditions. None of the commonly used housekeeping genes were present in the top 50 of the most stable expressed genes. In addition, using 2,543 diverse mouse gene array samples we were able to confirm the enhanced stability of the candidate novel housekeeping genes in another mammalian species. Therefore, the identified novel candidate housekeeping genes seem to be the most appropriate choice for normalizing gene expression data.

Multiple Testing Issues in Discriminating Compound-related Peaks and Chromatograms from High Frequency Noise, Spikes and Solvent-based Noise in LC-MS Data Sets

Statistical Applications in Genetics and Molecular Biology. 2007  |  Pubmed ID: 17910529

Liquid Chromatography--Mass Spectrometry (LC-MS) is a powerful method for sensitive detection and quantification of proteins and peptides in complex biological fluids like serum. LC-MS produces complex data sets, consisting of some hundreds of millions of data points per sample at a resolution of 0.1 amu in the m/z domain and 7000 data points in the time domain. However, the detection of the lower abundance proteins from this data is hampered by the presence of artefacts, such as high frequency noise and spikes. Moreover, not all of the tens of thousands of the chromatograms produced per sample are relevant for the pursuit of the biomarkers. Thus in analysing the LC-MS data, two critical pre-processing issues arise. Which of the thousands of the: 1. chromatograms per sample are relevant for the detection of the biomarkers?, and 2. signals per chromatogram are truly compound-related? Each of these issues involves assessing the significance (deviation from noise) of multiple observations and the issue of multiple comparisons arises. Current methods disregard the multiplicity and provide no concrete threshold for significance. However, with such procedures, the probability of one or more false-positives is high as the number of tests to be performed is large, and must be controlled. Realizing that the cut-offs for declaring a chromatogram (or a signal) to be compound-related can hugely influence which proteins are detected, it seems natural to define thresholds that are neither arbitrary nor subjective. We suggest the choice of thresholds guided by the critical aim of controlling the False Discovery Rate (FDR) in multiple hypotheses testing for significance over a large set of features produced per sample. This involves the use of the regression diagnostics to characterize the signals of a chromatogram (e.g. as outliers or influential) and to suggest suitable tests statistics for the multiple testing procedures (MTP) for discriminating noise and spikes from true signals. The role of the Generalized Linear Models (GLM) in this MTP is investigated. The method is applied to LC-MS datasets from trypsin-digested serum spiked with varying levels of horse heart cytochrome C (cytoc).

Combining Multiple Serum Tumor Markers Improves Detection of Stage I Epithelial Ovarian Cancer

Gynecologic Oncology. Dec, 2007  |  Pubmed ID: 17920110

Currently available tumor markers for ovarian cancer are still inadequate in both sensitivity and specificity to be used for population-based screening. Artificial neural network (ANN) as a modeling tool has demonstrated its ability to assimilate information from multiple sources and to detect subtle and complex patterns. In this paper, an ANN model was evaluated for its performance in detecting early stage epithelial ovarian cancer using multiple serum markers.

ESGO Statement on Cervical Cancer Vaccination

International Journal of Gynecological Cancer : Official Journal of the International Gynecological Cancer Society. Nov-Dec, 2007  |  Pubmed ID: 17997794

Chip-LC-MS for Label-free Profiling of Human Serum

Electrophoresis. Dec, 2007  |  Pubmed ID: 18041038

The discovery of biomarkers in easily accessible body fluids such as serum is one of the most challenging topics in proteomics requiring highly efficient separation and detection methodologies. Here, we present the application of a microfluidics-based LC-MS system (chip-LC-MS) to the label-free profiling of immunodepleted, trypsin-digested serum in comparison to conventional capillary LC-MS (cap-LC-MS). Both systems proved to have a repeatability of approximately 20% RSD for peak area, all sample preparation steps included, while repeatability of the LC-MS part by itself was less than 10% RSD for the chip-LC-MS system. Importantly, the chip-LC-MS system had a two times higher resolution in the LC dimension and resulted in a lower average charge state of the tryptic peptide ions generated in the ESI interface when compared to cap-LC-MS while requiring approximately 30 times less (~5 pmol) sample. In order to characterize both systems for their capability to find discriminating peptides in trypsin-digested serum samples, five out of ten individually prepared, identical sera were spiked with horse heart cytochrome c. A comprehensive data processing methodology was applied including 2-D smoothing, resolution reduction, peak picking, time alignment, and matching of the individual peak lists to create an aligned peak matrix amenable for statistical analysis. Statistical analysis by supervised classification and variable selection showed that both LC-MS systems could discriminate the two sample groups. However, the chip-LC-MS system allowed to assign 55% of the overall signal to selected peaks against 32% for the cap-LC-MS system.

Sentinel Node Dissection is Safe in the Treatment of Early-stage Vulvar Cancer

Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Feb, 2008  |  Pubmed ID: 18281661

To investigate the safety and clinical utility of the sentinel node procedure in early-stage vulvar cancer patients.

A New Perspective on Transcriptional System Regulation (TSR): Towards TSR Profiling

PloS One. 2008  |  Pubmed ID: 18297136

It has been hypothesized that the net expression of a gene is determined by the combined effects of various transcriptional system regulators (TSRs). However, characterizing the complexity of regulation of the transcriptome is a major challenge. Principal component analysis on 17,550 heterogeneous human microarray experiments revealed that 50 orthogonal factors (hereafter called TSRs) are able to capture 64% of the variability in expression in a wide range of experimental conditions and tissues. We identified gene clusters controlled in the same direction and show that gene expression can be conceptualized as a process influenced by a fairly limited set of TSRs. Furthermore, TSRs can be linked to biological functions, as we demonstrate a strong relation between TSR-related gene clusters and biological functionality as well as cellular localization, i.e. gene products of similarly regulated genes by a specific TSR are located in identical parts of a cell. Using 3,934 diverse mouse microarray experiments we found striking similarities in transcriptional system regulation between human and mouse. Our results give biological insights into regulation of the cellular transcriptome and provide a tool to characterize expression profiles with highly reliable TSRs instead of thousands of individual genes, leading to a >500-fold reduction of complexity with just 50 TSRs. This might open new avenues for those performing gene expression profiling studies.

The Effect of Pre-existing Immunity on the Capacity of Influenza Virosomes to Induce Cytotoxic T Lymphocyte Activity

Vaccine. May, 2008  |  Pubmed ID: 18400343

Protein antigens encapsulated in virosomes generated from influenza virus can induce antigen-specific cytotoxic T lymphocyte (CTL) responses. In the present study we determined, in a murine model system, whether pre-existing immunity against influenza virus hampers the induction of a CTL response. CTL induction was only slightly reduced by pre-injection of influenza virus-specific antibodies or pre-exposure to influenza virus. Both pretreatments resulted in the same level of reduction, suggesting that virus-specific antibodies rather than T cell responses account for the reduction. Furthermore, a booster immunization enhanced CTL activation, indicating that virosome-specific immunity induced by a prime immunization does not hamper the booster effect. In conclusion, CTL induction against virosome-encapsulated protein antigens is not significantly inhibited by pre-existing humoral or cellular immunity against influenza virus.

Genome-wide Promoter Analysis Uncovers Portions of the Cancer Methylome

Cancer Research. Apr, 2008  |  Pubmed ID: 18413733

DNA methylation has a role in mediating epigenetic silencing of CpG island genes in cancer and other diseases. Identification of all gene promoters methylated in cancer cells "the cancer methylome" would greatly advance our understanding of gene regulatory networks in tumorigenesis. We previously described a new method of identifying methylated tumor suppressor genes based on pharmacologic unmasking of the promoter region and detection of re-expression on microarray analysis. In this study, we modified and greatly improved the selection of candidates based on new promoter structure algorithm and microarray data generated from 20 cancer cell lines of 5 major cancer types. We identified a set of 200 candidate genes that cluster throughout the genome of which 25 were previously reported as harboring cancer-specific promoter methylation. The remaining 175 genes were tested for promoter methylation by bisulfite sequencing or methylation-specific PCR (MSP). Eighty-two of 175 (47%) genes were found to be methylated in cell lines, and 53 of these 82 genes (65%) were methylated in primary tumor tissues. From these 53 genes, cancer-specific methylation was identified in 28 genes (28 of 53; 53%). Furthermore, we tested 8 of the 28 newly identified cancer-specific methylated genes with quantitative MSP in a panel of 300 primary tumors representing 13 types of cancer. We found cancer-specific methylation of at least one gene with high frequency in all cancer types. Identification of a large number of genes with cancer-specific methylation provides new targets for diagnostic and therapeutic intervention, and opens fertile avenues for basic research in tumor biology.

A Robust Ex Vivo Model for Evaluation of Induction of Apoptosis by RhTRAIL in Combination with Proteasome Inhibitor MG132 in Human Premalignant Cervical Explants

International Journal of Cancer. Journal International Du Cancer. Sep, 2008  |  Pubmed ID: 18567003

Development of medical therapies for high-grade cervical intraepithelial neoplasia (CIN II/III) is hampered by the lack of CIN II/III cell lines. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis upon binding to its receptors DR4 or DR5. Proteasome inhibition by MG132 sensitized cervical cancer cell lines to recombinant human (rh)TRAIL. In our study, we aimed to develop an ex vivo model for CIN II/III and to investigate the apoptosis-inducing effect of rhTRAIL and/or MG132 in cervical explants from CIN II/III patients. A short-term ex vivo culture system was optimized for cervical biopsies, in which explants from normal cervix and CIN II/III lesions were exposed to either rhTRAIL (1 microg/ml), MG132 (5 microM) or the combination and compared to untreated explants for apoptosis induction. Normal cervix (n = 90) and CIN II/III (n = 24) explants could be reproducibly put in culture and kept viable for up to 7 days using a transwell membrane system. CIN II/III explants (n = 5) were highly sensitive to rhTRAIL plus MG132 (mean % apoptosis: 91 +/- 5) compared to normal cervix (n = 10) treated with rhTRAIL plus MG132 (mean % apoptosis: 24 +/- 10, p < 0.0001), while monotherapy with either rhTRAIL, MG132 or medium resulted in a mean % apoptosis <10 in both CIN II/III and normal cervix. Our ex vivo model system allows preclinical evaluation of (topical) medical therapies for CIN II/III. A strong synergistic apoptosis-inducing effect of the combination of rhTRAIL and MG132, especially in CIN II/III lesions indicates that rhTRAIL combined with proteasome inhibitors deserves exploration as medical treatment for CIN II/III.

Survival of Ovarian Cancer Patients Overexpressing the Tumour Antigen P53 is Diminished in Case of MHC Class I Down-regulation

Gynecologic Oncology. Sep, 2008  |  Pubmed ID: 18571704

The adaptive immune system seems to play an essential role in the natural course of ovarian cancer. Aim of this study was to establish whether disease-specific survival for patients expressing the tumour antigen p53 is influenced by MHC class I expression or the presence of p53 autoantibodies (p53-Aab).

Targeting Pro-apoptotic Trail Receptors Sensitizes HeLa Cervical Cancer Cells to Irradiation-induced Apoptosis

International Journal of Radiation Oncology, Biology, Physics. Oct, 2008  |  Pubmed ID: 18793956

To investigate the potential of irradiation in combination with drugs targeting the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor (DR)4 and DR5 and their mechanism of action in a cervical cancer cell line.

Distinguishing Benign and Malignant Pelvic Masses: the Value of Different Diagnostic Methods in Everyday Clinical Practice

European Journal of Obstetrics, Gynecology, and Reproductive Biology. Jan, 2008  |  Pubmed ID: 17118522

To optimize referral to specialized gynaecologists for surgical treatment of ovarian cancer by improving preoperative discrimination between benign and malignant pelvic tumours.

Survival-related Profile, Pathways, and Transcription Factors in Ovarian Cancer

PLoS Medicine. Feb, 2009  |  Pubmed ID: 19192944

Ovarian cancer has a poor prognosis due to advanced stage at presentation and either intrinsic or acquired resistance to classic cytotoxic drugs such as platinum and taxoids. Recent large clinical trials with different combinations and sequences of classic cytotoxic drugs indicate that further significant improvement in prognosis by this type of drugs is not to be expected. Currently a large number of drugs, targeting dysregulated molecular pathways in cancer cells have been developed and are introduced in the clinic. A major challenge is to identify those patients who will benefit from drugs targeting these specific dysregulated pathways.The aims of our study were (1) to develop a gene expression profile associated with overall survival in advanced stage serous ovarian cancer, (2) to assess the association of pathways and transcription factors with overall survival, and (3) to validate our identified profile and pathways/transcription factors in an independent set of ovarian cancers.

[Primary Surgery by a Gynecological Oncologist Improves the Prognosis in Patients with Ovarian Carcinoma]

Nederlands Tijdschrift Voor Geneeskunde. Jan, 2009  |  Pubmed ID: 19198207

[The Limitations of Observational Research]

Nederlands Tijdschrift Voor Geneeskunde. Jan, 2009  |  Pubmed ID: 19235351

Down-regulation of Proteasomal Subunit MB1 is an Independent Predictor of Improved Survival in Ovarian Cancer

Gynecologic Oncology. May, 2009  |  Pubmed ID: 19243813

To investigate the expression and to determine the prognostic impact of components of the antigen processing and presentation pathway (APPP) in ovarian cancer.

Enhanced Antitumor Efficacy of a DR5-specific TRAIL Variant over Recombinant Human TRAIL in a Bioluminescent Ovarian Cancer Xenograft Model

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Mar, 2009  |  Pubmed ID: 19276284

Recombinant human tumor necrosis factor-related apoptosis-inducing ligand (rhTRAIL) is clinically evaluated as novel anticancer drug. rhTRAIL-DR5, a rhTRAIL variant that specifically binds to DR5 receptor, has recently been developed. We investigated whether rhTRAIL-DR5 is more efficient than rhTRAIL in combination with cisplatin in DR5-expressing human A2780 ovarian cancer cells. Design: Effect of cisplatin alone or in combination with rhTRAIL or rhTRAIL-DR5 on DR5 surface expression, apoptosis, and cell survival of A2780 was measured. Biodistribution analysis was done in mice with (125)I-rhTRAIL administered intravenously versus intraperitoneally. Antitumor efficacy of rhTRAIL-DR5 versus rhTRAIL was determined in an intraperitoneally growing bioluminescent A2780 xenograft model.

Sentinel Lymph Node Biopsy in Patients with Gynecologic Cancers Expert Panel Statement from the International Sentinel Node Society Meeting, February 21, 2008

Gynecologic Oncology. Aug, 2009  |  Pubmed ID: 19457548

An expert panel was formed for the 6th biennial International Sentinel Node Society to review the status of sentinel lymph node biopsy (SLNB) in gynecologic oncology. This paper presents the opinion of the experts who participated regarding indications for SLNB, technical considerations, and directions for future investigation.

Aromatase, Cyclooxygenase 2, HER-2/neu, and P53 As Prognostic Factors in Endometrioid Endometrial Cancer

International Journal of Gynecological Cancer : Official Journal of the International Gynecological Cancer Society. May, 2009  |  Pubmed ID: 19509570

The prognostic value of aromatase, cyclooxygenase 2 (COX-2), HER-2/neu, and p53 expression was determined in endometrioid endometrial cancer. Tissue microarrays were constructed comprising samples from 315 endometrioid endometrial cancer patients. Expression of aromatase, COX-2, HER-2/neu, and p53 was determined by immunostaining and related to classical clinicohistopathologic parameters, in addition to recurrence of disease and survival. Median follow-up time for all patients was 5.0 years. Patients were classified as Fédération Internationale de Gynécologie Obstétrique stage I (59.0%), stage II (17.1%), stage III (19.4%), and stage IV (4.1%). Sixty-five patients (20.6%) developed recurrent disease, and 38 (12.1%) died because of endometrial cancer. Aromatase, COX-2, HER-2/neu, and p53 expression was observed in 133 (42.2%), 107 (34.0%), 17 (5.4%), and 21 (6.7%) tumor cases, respectively. Aromatase expression in tumor cells was related to aromatase expression in stromal cells (P < 0.0001) and to HER-2/neu expression in tumor cells (P = 0.019). Aromatase expression in tumor as well as stromal cells was related to a low stage of disease (P = 0.02 and P = 0.001, respectively), whereas aromatase expression in stromal cells was also related to a low tumor grade (P = 0.021). P53 expression was related to a high stage and a high grade (P = 0.006 and P < 0.0001, respectively). In multivariate analysis, p53 overexpression was independently related to death because of the disease (P = 0.043; odds ratio 3.0; 95% confidence interval, 1.0-8.7). For COX-2, HER-2/neu, and aromatase, no relation with any other histopathologic parameter or survival was found. In conclusion, aromatase and p53 expression are related to tumor grade and stage of disease, whereas p53 is an independent prognostic factor in endometrioid endometrial cancer.

The Role of Sentinel Node Biopsy in Gynecological Cancer: a Review

Current Opinion in Oncology. Sep, 2009  |  Pubmed ID: 19593136

In early-stage vulvar, cervical and endometrial cancer, lymph node status is the most important prognostic factor. Surgical treatment is aimed at removing the primary tumor and adequately staging the regional lymph nodes. As morbidity of regional lymphadenectomy is high, sentinel node biopsy is a technique with potential for adequate staging with less treatment-related morbidity. This manuscript reviews its current role in vulvar, cervical and endometrial cancer.

Immunization with a P53 Synthetic Long Peptide Vaccine Induces P53-specific Immune Responses in Ovarian Cancer Patients, a Phase II Trial

International Journal of Cancer. Journal International Du Cancer. Nov, 2009  |  Pubmed ID: 19621448

The prognosis of ovarian cancer, the primary cause of death from gynecological malignancies, has only modestly improved over the last decades. Immunotherapy is one of the new treatment modalities explored for this disease. To investigate safety, tolerability, immunogenicity and obtain an impression of clinical activity of a p53 synthetic long peptide (p53-SLP) vaccine, twenty patients with recurrent elevation of CA-125 were included, eighteen of whom were immunized 4 times with 10 overlapping p53-SLP in Montanide ISA51. The first 5 patients were extensively monitored for toxicity, but showed no > or = grade 3 toxicity, thus accrual was continued. Overall, toxicity was limited to grade 1 and 2, mostly locoregional, inflammatory reactions. IFN-gamma producing p53-specific T-cell responses were induced in all patients who received all 4 immunizations as measured by IFN-gamma ELISPOT. An IFN-gamma secretion assay showed that vaccine-induced p53-specific T-cells were CD4(+), produced both Th1 and Th2 cytokines as analyzed by cytokine bead array. Notably, Th2 cytokines dominated the p53-specific response. P53-specific T-cells were present in a biopsy of the last immunization site of at least 9/17 (53%) patients, reflecting the migratory capacity of p53-specific T-cells. As best clinical response, stable disease evaluated by CA-125 levels and CT-scans, was observed in 2/20 (10%) patients, but no relationship was found with vaccine-induced immunity. This study shows that the p53-SLP vaccine is safe, well tolerated and induces p53-specific T-cell responses in ovarian cancer patients. Upcoming trials will focus on improving T helper-1 polarization and clinical efficacy.

TP53 Codon 72 Polymorphism and Cervical Cancer: a Pooled Analysis of Individual Data from 49 Studies

The Lancet Oncology. Aug, 2009  |  Pubmed ID: 19625214

Cervical cancer is caused primarily by human papillomaviruses (HPV). The polymorphism rs1042522 at codon 72 of the TP53 tumour-suppressor gene has been investigated as a genetic cofactor. More than 80 studies were done between 1998 and 2006, after it was initially reported that women who are homozygous for the arginine allele had a risk for cervical cancer seven times higher than women who were heterozygous for the allele. However, results have been inconsistent. Here we analyse pooled data from 49 studies to determine whether there is an association between TP53 codon 72 polymorphism and cervical cancer.

The Prognostic Value of TRAIL and Its Death Receptors in Cervical Cancer

International Journal of Radiation Oncology, Biology, Physics. Sep, 2009  |  Pubmed ID: 19695437

Preclinical data indicate a synergistic effect on apoptosis between irradiation and recombinant human (rh) tumor necrosis factor-related apoptosis inducing ligand (TRAIL), making the TRAIL death receptors (DR) interesting drug targets. The aim of our study was to analyze the expression of DR4, DR5, and TRAIL in cervical cancer and to determine their predictive and prognostic value.

Identification of Candidate Epigenetic Biomarkers for Ovarian Cancer Detection

Oncology Reports. Oct, 2009  |  Pubmed ID: 19724865

Ovarian cancer ranks the most lethal among gynecologic neoplasms in women. To develop potential biomarkers for diagnosis, we have identified five novel genes (CYP39A1, GTF2A1, FOXD4L4, EBP, and HAAO) that are hypermethylated in ovarian tumors, compared with the non-malignant normal ovarian surface epithelia, using the quantitative methylation-specific polymerase chain reactions. Interestingly enough, multivariate Cox regression analysis has identified hypermethylation of CYP39A1 correlated with an increase rate of relapsing (P=0.032, hazard ratio >1). Concordant hypermethylation in at least three loci was observed in 50 out of 55 (91%) of ovarian tumors examined. The test sensitivity and specificity were assessed to be 96 and 67% for CYP39A1; 95 and 88% for GTF2A1; 93 and 67% for FOXD4L4; 81 and 67% for EBP; 89 and 82% for HAAO, respectively. Our data have identified, for the first time, GTF2A1 alone, or GTF2A1 plus HAAO are excellent candidate biomarkers for detecting this disease. Moreover, the known functions of these gene products further implicate dysregulated transcriptional control, cholesterol metabolism, or synthesis of quinolinic acids, may play important roles in attributing to ovarian neoplasm. Molecular therapies, by reversing the aberrant epigenomes using inhibitory agents or by abrogating the upstream signaling pathways that convey the epigenomic perturbations, may be developed into promising treatment regimens.

[New Strategies for Screening Cervical Cancer; Response to the Dutch College of General Practitioner's Practice Guideline 'Prevention and Early Diagnosis of Cervical Cancer']

Nederlands Tijdschrift Voor Geneeskunde. 2009  |  Pubmed ID: 19785834

The publication of a practice guideline for primary and secondary prevention of cervical cancer by the Dutch College of General Practitioners was appropriate since the vaccination against two high-risk human papillomaviruses (hr-HPV 16 and 18) was recently included in the National Immunization Programme of the Netherlands. Despite vaccination, population-based screening for cervical neoplasia needs to be continued. Moreover, it will take over a decade before vaccination will affect the prevalence of cervical neoplasia. Short term improvement of prevention of cervical neoplasia should therefore focus on screening technology and strategy. Molecular tests based on detection of hr-HPV or DNA changes occurring early in cervical carcinogenesis might improve the efficacy of the screening, which in the Netherlands is currently done by Pap smear. In addition, the high number of women not responding on an invitation for screening might be reduced by a self-sampling approach.

Methylation Markers for CCNA1 and C13ORF18 Are Strongly Associated with High-grade Cervical Intraepithelial Neoplasia and Cervical Cancer in Cervical Scrapings

Cancer Epidemiology, Biomarkers & Prevention : a Publication of the American Association for Cancer Research, Cosponsored by the American Society of Preventive Oncology. Nov, 2009  |  Pubmed ID: 19843677

Recently, we reported 13 possible cervical cancer-specific methylated biomarkers identified by pharmacologic unmasking microarray in combination with large-genome computational screening. The aim of the present study was to perform an in-depth analysis of the methylation patterns of these 13 candidate genes in cervical neoplasia and to determine their diagnostic relevance.

Expression of Epidermal Growth Factor Receptor (EGFR) and Activated EGFR Predict Poor Response to (chemo)radiation and Survival in Cervical Cancer

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Dec, 2009  |  Pubmed ID: 19920104

Activation of the epidermal growth factor receptor (EGFR) signaling pathway has been reported to induce resistance to (chemo)radiation in cancers, such as head and neck cancer, whereas EGFR-targeted agents in combination with (chemo)radiation seem to improve treatment efficacy. The aim of this study was to determine the relation between proteins involved in the EGFR pathway and response to (chemo)radiation and survival in a large, well-documented series of cervical cancer patients.

Prognostic Significance of Tumor-infiltrating T-lymphocytes in Primary and Metastatic Lesions of Advanced Stage Ovarian Cancer

Cancer Immunology, Immunotherapy : CII. Mar, 2009  |  Pubmed ID: 18791714

Ovarian cancer patients with intra-tumoral CD3(+) T-lymphocytes in primary tumor tissue have a better prognosis. This study aims to analyze the presence and relative influence of three important T-lymphocyte subsets, tumor-infiltrating CD8(+) cytotoxic T-lymphocytes (CTL), CD45R0(+) memory T-lymphocytes, and FoxP3(+) regulatory T-lymphocytes (Treg), in primary tumor tissue and omental metastases of patients with ovarian cancer.

Multimodality Treatment Warranted for Ovarian Cancer: Immunotherapy, a Prerequisite to Improve Prognosis for This Vicious Disease

Immunotherapy. Mar, 2009  |  Pubmed ID: 20635935

Potential Target Antigens for a Universal Vaccine in Epithelial Ovarian Cancer

Clinical & Developmental Immunology. 2010  |  Pubmed ID: 20885926

The prognosis of epithelial ovarian cancer (EOC), the primary cause of death from gynaecological malignancies, has only modestly improved over the last decades. Immunotherapeutic treatment using a cocktail of antigens has been proposed as a "universal" vaccine strategy. We determined the expression of tumor antigens in the context of MHC class I expression in 270 primary tumor samples using tissue microarray. Expression of tumor antigens p53, SP17, survivin, WT1, and NY-ESO-1 was observed in 120 (48.0%), 173 (68.9%), 208 (90.0%), 129 (56.3%), and 27 (11.0%) of 270 tumor specimens, respectively. In 93.2% of EOC, at least one of the investigated tumor antigens was (over)expressed. Expression of MHC class I was observed in 78.1% of EOC. In 3 out 4 primary tumors, (over)expression of a tumor antigen combined with MHC class I was observed. These results indicate that a multiepitope vaccine, comprising these antigens, could serve as a universal therapeutic vaccine for the vast majority of ovarian cancer patients.

Implementing an Advanced Laparoscopic Procedure by Monitoring with a Visiting Surgeon

Journal of Minimally Invasive Gynecology. Nov-Dec, 2010  |  Pubmed ID: 20955987

To investigate the feasibility of safely implementing a total laparoscopic hysterectomy (LH) in established gynecologists' practices with on-site coaching and monitoring of the learning curve by an experienced visiting surgeon.

The Epidermal Growth Factor Receptor Pathway in Relation to Pelvic Lymph Node Metastasis and Survival in Early-stage Cervical Cancer

Human Pathology. Dec, 2010  |  Pubmed ID: 21078436

The objective of this study is to correlate the expression of epidermal growth factor receptor (EGFR) components with clinical behavior of early-stage cervical cancer. Tissue samples of 336 consecutive Federation of International Gynecologists and Obstetricians stage IB-IIA cervical cancer patients all treated primarily by radical surgery were collected. Clinicopathologic and follow-up data were prospectively obtained during standard treatment and follow-up. As representatives for the EGFR pathway, expression of EGFR, pEGFR, PTEN, pAKT, and pERK was assessed by immunohistochemistry on tissue microarrays. Positive immunostaining was observed for EGFR in 32.1%, for pEGFR in 21.0%, for PTEN in 38.3%, for pAKT in 5.3%, and for pERK in 4.3% of tumor samples. Positive EGFR immunostaining was associated with squamous cell carcinoma of the cervix (odds ratio [OR], 7.41; 95% confidence interval [CI], 3.38-16.23, P < .001), negative pEGFR immunostaining with poor differentiation (OR, 0.39; 95% CI, 0.20-0.73, P = .004), and negative PTEN immunostaining with metastatic pelvic lymph nodes (OR, 0.51; 95% CI, 0.30-0.90, P = .019). In multivariate analysis, only pelvic lymph node metastasis (hazard ratio, 6.11; 95% CI, 3.46-10.77, P < .001) and poor differentiation (hazard ratio, 1.91; 95% CI, 1.12-3.26, P = .018) were related to disease-specific survival. In early-stage cervical cancer, loss of PTEN expression is associated with pelvic lymph node metastasis, suggesting PTEN to be one of the tumor suppressor genes affecting pelvic lymph node metastasis. However, expression of EGFR pathway components does not appear to have prognostic impact in surgically treated early-stage cervical cancer.

The Extrinsic Apoptosis Pathway and Its Prognostic Impact in Ovarian Cancer

Gynecologic Oncology. Mar, 2010  |  Pubmed ID: 19959214

Death ligand FasL, its agonistic receptor Fas, tumor necrosis factor related apoptosis inducing ligand (TRAIL) and its agonistic death receptors DR4 and DR5 are implied in carcinogenesis, tumor immune surveillance and response to chemotherapy. TRAIL receptor agonists are evaluated as anti-cancer agents. This study aimed to relate expression of death ligands/receptors and downstream initiator caspase 8 and its anti-apoptotic homologue FLICE like inhibitory protein (c-FLIP) in ovarian cancers to chemotherapy response and survival.

Update on the Sentinel Lymph Node Procedure in Vulvar Cancer

Expert Review of Anticancer Therapy. Jan, 2010  |  Pubmed ID: 20014886

Currently, standard treatment for early-stage vulvar cancer typically includes wide local excision of the primary tumor and inguinofemoral lymphadenectomy. The morbidity of this treatment is high. The sentinel lymph node (SLN) procedure provides us with a technique for determining the status of the regional lymph nodes with less treatment-related morbidity. Recently, a large multicenter observational study provided level 3 evidence indicating that it appears safe to omit inguinofemoral lymphadenectomy in case of a negative SLN. This review focuses on the different aspects of the SLN procedure in vulvar cancer.

Gene Promoter Methylation Patterns Throughout the Process of Cervical Carcinogenesis

Cellular Oncology : the Official Journal of the International Society for Cellular Oncology. 2010  |  Pubmed ID: 20208141

To determine methylation status of nine genes, previously described to be frequently methylated in cervical cancer, in squamous intraepithelial lesions (SIL).

European Network of Gynaecological Oncological Trial Groups' Requirements for Trials Between Academic Groups and Pharmaceutical Companies

International Journal of Gynecological Cancer : Official Journal of the International Gynecological Cancer Society. Apr, 2010  |  Pubmed ID: 20375816

Routine Evaluation and Treatment of Unexplained Menorrhagia: Do We Consider Haemostatic Disorders?

European Journal of Obstetrics, Gynecology, and Reproductive Biology. Oct, 2010  |  Pubmed ID: 20576344

Unexplained menorrhagia can be caused by underlying bleeding disorders. The aim of this study was to investigate the current work-up of menorrhagia in routine gynaecological practice, with a special interest in haemostatic evaluation. Secondly, we investigated the outcome of individualized treatment in our centre.

Current Controversies in the Management of Patients with Early-stage Vulvar Cancer

Current Opinion in Oncology. Sep, 2010  |  Pubmed ID: 20616719

The purpose of this review is to outline current controversies in management of early-stage vulvar cancer. The main focus will be on the procedures for assessing the sentinel node and the treatment of those with evidence of metastatic involvement.

The Prognostic Role of Classical and Nonclassical MHC Class I Expression in Endometrial Cancer

International Journal of Cancer. Journal International Du Cancer. Mar, 2010  |  Pubmed ID: 19728333

The aim of this study was to investigate classical MHC class I and nonclassical MHC (human leukocyte antigen-G [HLA-G]) expression in a large cohort of patients with endometrial cancer, to determine the prognostic value of these cell surface markers and their relation with clinicopathological variables. Tissue microarrays containing epithelial endometrial carcinoma tissue from 554 patients were stained for classical and nonclassical MHC class I using the following monoclonal antibodies: 4H84 (anti-HLA-G), beta2-m (anti-beta-2-microglobulin) and HC-10 (MHC class I antigen heavy chain). Expression data were linked to known clinicopathological characteristics and survival. HLA-G upregulation and MHC class I downregulation in neoplastic cells was observed in 40% and 48%, respectively. Nonendometrioid tumor type, advanced stage disease (FIGO stage > or = II) and poorly or undifferentiated tumors were associated with MHC class I downregulation. Absence of HLA-G expression was independently associated with MHC class I downregulation. In univariate analysis, MHC class I downregulation was a predictor of worse disease-specific survival. Prognostic unfavorable tumor characteristics were correlated with downregulation of MHC class I expression in endometrial cancer cells. Furthermore, downregulated MHC class I has a negative impact on disease-specific survival, observed in a large cohort of patients with endometrial cancer. As there seems to be a relation between classical and nonclassical MHC class I molecules (HLA-G), further research is warranted to unravel this regulatory mechanism.

Loss of SerpinA5 Protein Expression is Associated with Advanced-stage Serous Ovarian Tumors

Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc. Mar, 2011  |  Pubmed ID: 21102419

Epithelial ovarian cancer, the most lethal neoplasm of the female genital tract, is usually diagnosed at an advanced stage as obvious symptoms are absent at early stages. This disease is believed to originate from malignant transformation of the ovarian surface epithelium or fallopian tube. Histologically, several subtypes are being recognized, with serous histology accounting for the majority of cases. Serous tumors include serous borderline tumors and serous carcinomas. A better understanding of the tumor biology and molecular mechanisms involved in these tumors is needed, as both patient management and prognosis differ substantially. Previous microarray analysis identified SerpinA5, a uPA inhibitor, as key regulator for indolent borderline behavior. As carcinomas are characterized by loss of SerpinA5 mRNA expression, we hypothesized that SerpinA5 protein expression is reduced or lost in carcinomas when compared with borderline tumors. We performed SerpinA5 immunohistochemical staining on 32 serous borderline tumors, 187 primary serous carcinomas and 62 serous omental metastases. Reduced or absent SerpinA5 protein staining was observed in carcinomas when compared with borderline tumors (P<0.001). SerpinA5 protein expression was significantly lowered in the omental metastases (P<0.001) when compared with the matching primary carcinoma. Interestingly, SerpinA5 protein expression was reduced in advanced-stage borderline tumors, often characterized by micropapillary growth and/or microinvasion, when compared with early-stage borderline tumors (P=0.015). In conclusion, SerpinA5 expression is significantly reduced in advanced-stage serous borderline tumors and serous carcinomas when compared with the early-stage counterparts, and reduction of expression is linked to more aggressive features of borderline tumors.

Prognostic Cell Biological Markers in Cervical Cancer Patients Primarily Treated with (chemo)radiation: a Systematic Review

International Journal of Radiation Oncology, Biology, Physics. Feb, 2011  |  Pubmed ID: 21195874

The aim of this study was to systematically review the prognostic and predictive significance of cell biological markers in cervical cancer patients primarily treated with (chemo)radiation. A PubMed, Embase, and Cochrane literature search was performed. Studies describing a relation between a cell biological marker and survival in ≥50 cervical cancer patients primarily treated with (chemo)radiation were selected. Study quality was assessed, and studies with a quality score of 4 or lower were excluded. Cell biological markers were clustered on biological function, and the prognostic and predictive significance of these markers was described. In total, 42 studies concerning 82 cell biological markers were included in this systematic review. In addition to cyclooxygenase-2 (COX-2) and serum squamous cell carcinoma antigen (SCC-ag) levels, markers associated with poor prognosis were involved in epidermal growth factor receptor (EGFR) signaling (EGFR and C-erbB-2) and in angiogenesis and hypoxia (carbonic anhydrase 9 and hypoxia-inducible factor-1α). Epidermal growth factor receptor and C-erbB-2 were also associated with poor response to (chemo)radiation. In conclusion, EGFR signaling is associated with poor prognosis and response to therapy in cervical cancer patients primarily treated with (chemo)radiation, whereas markers involved in angiogenesis and hypoxia, COX-2, and serum SCC-ag levels are associated with a poor prognosis. Therefore, targeting these pathways in combination with chemoradiation may improve survival in advanced-stage cervical cancer patients.

Involvement of the TGF-beta and Beta-catenin Pathways in Pelvic Lymph Node Metastasis in Early-stage Cervical Cancer

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Mar, 2011  |  Pubmed ID: 21385933

Presence of pelvic lymph node metastases is the main prognostic factor in early-stage cervical cancer patients, primarily treated with surgery. Aim of this study was to identify cellular tumor pathways associated with pelvic lymph node metastasis in early-stage cervical cancer.

Intraoperative Imaging in Ovarian Cancer: Fact or Fiction?

Molecular Imaging. Aug, 2011  |  Pubmed ID: 21521557

Tumor-targeted fluorescence imaging for cancer diagnosis and treatment is an evolving field of research that is on the verge of clinical implementation. As each tumor has its unique biologic profile, selection of the most promising targets is essential. In this review, we focus on target finding in ovarian cancer, a disease in which fluorescence imaging may be of value in both adequate staging and in improving cytoreductive efforts, and as such may have a beneficial effect on prognosis. Thus far, tumor-targeted imaging for ovarian cancer has been applied only in animal models. For clinical implementation, the five most prominent targets were identified: folate receptor α, vascular endothelial growth factor, epidermal growth factor receptor, chemokine receptor 4, and matrix metalloproteinase. These targets were selected based on expression rates in ovarian cancer, availability of an antibody or substrate aimed at the target approved by the Food and Drug Administration, and the likelihood of translation to human use. The purpose of this review is to present requirements for intraoperative imaging and to discuss possible tumor-specific targets for ovarian cancer, prioritizing for targets with substrates ready for introduction into the clinic.

Laparoscopic Hysterectomy is Preferred over Laparotomy in Early Endometrial Cancer Patients, However Not Cost Effective in the Very Obese

European Journal of Cancer (Oxford, England : 1990). Sep, 2011  |  Pubmed ID: 21636268

Total laparoscopic hysterectomy (TLH) is safe and cost effective in early stage endometrial cancer when compared to total abdominal hysterectomy (TAH). In non-randomised data it is often hypothesised that older and obese patients benefit most from TLH. Aim of this study is to analyse whether data support this assumption to advice patients, clinicians and policy makers.

Differentiated Vulvar Intraepithelial Neoplasia is Often Found in Lesions, Previously Diagnosed As Lichen Sclerosus, Which Have Progressed to Vulvar Squamous Cell Carcinoma

Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc. Feb, 2011  |  Pubmed ID: 21057461

Lichen sclerosus is considered to be the precursor lesion of vulvar squamous cell carcinoma, of which only 2-5% progress to squamous cell carcinoma. Differentiated vulvar intraepithelial neoplasia (VIN) has been proposed to be the direct precursor lesion, but this is a recently recognized, and a difficult to diagnose, entity, which may easily be mistaken for a benign dermatosis. The aim of this study was to test the hypothesis that of all lesions that have been diagnosed as lichen sclerosus in the past, a part might currently be diagnosed as differentiated VIN, and to identify histopathological differences between lichen sclerosus lesions with and without progression to vulvar squamous cell carcinoma. All lichen sclerosus slides were revised by two expert gynecopathologists and histopathological characteristics were documented. After revision of lichen sclerosus biopsies without progression (n = 61), 58 were reclassified as lichen sclerosus. Revision of lichen sclerosus biopsies with progression yielded concordant diagnoses in 18 of 60 cases (30%). Of 60 lesions, 25 (42%) were reclassified as differentiated VIN. The median time from differentiated VIN to vulvar squamous cell carcinoma was shorter (28 months) than that from lichen sclerosus to vulvar squamous cell carcinoma (84 months) (P < 0.001). Lichen sclerosus that progressed to squamous cell carcinoma, but did not meet the criteria for differentiated VIN, more often showed parakeratosis (P = 0.004), dyskeratosis (P < 0.001), hyperplasia (P = 0.048) and basal cellular atypia (P = 0.009) compared with lichen sclerosus without progression. In conclusion, differentiated VIN diagnosis has been frequently missed and is associated with rapid progression to squamous cell carcinoma. Patients with lichen sclerosus with dyskeratosis and parakeratosis, hyperplasia and/or basal cellular atypia should be kept under close surveillance as these lesions also tend to progress to squamous cell carcinoma.

Heterologous Prime-boost Immunizations with a Virosomal and an Alphavirus Replicon Vaccine

Molecular Pharmaceutics. Feb, 2011  |  Pubmed ID: 20825215

Heterologous prime-boost immunization strategies in general establish higher frequencies of antigen-specific T lymphocytes than homologous prime-boost protocols or single immunizations. We developed virosomes and recombinant Semliki Forest virus (rSFV) as antigen delivery systems, each capable of inducing strong CTL responses in homologous prime-boost protocols. Here, we demonstrate that a heterologous prime-boost with recombinant Semliki Forest virus (rSFV) encoding a fusion protein of E6 and E7 of human papillomavirus (HPV) type 16 and virosomes containing the HPV16 E7 protein resulted in higher numbers of antigen-specific CTL in mice than homologous protocols. Evasion of vector-specific immunity appeared to play a role in establishing these high frequencies, as coinduction of vector-specific responses during the prime immunization reduced the frequency of antigen-specific CTL after a heterologous booster. However, the high numbers of CTL initially primed by the heterologous protocols did not correlate with enhanced responsiveness to in vitro antigenic stimulation, nor in improved cytolytic activity or antitumor responses in vivo compared to a homologous protocol with rSFV. This lack of correlation could not be explained by changes in numbers of regulatory T cells. However, we observed differences in the frequencies of T cell subsets within the E7-specific CD8(+) T cell population, e.g. higher frequencies of central memory T cells upon homologous immunizations compared to heterologous immunizations. The induction of central memory T cells is crucial for a cancer vaccine as these cells are known to rapidly expand upon recall stimulation. This study demonstrates that the strongly increased number of antigen-specific CTL as induced by heterologous prime-boost immunizations, often used as a proof for the enhanced efficacy of such regimes, does not necessarily equal superior functional antitumor responses.

Intraoperative Multispectral Fluorescence Imaging for the Detection of the Sentinel Lymph Node in Cervical Cancer: a Novel Concept

Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging. Oct, 2011  |  Pubmed ID: 20835767

Real-time intraoperative near-infrared fluorescence (NIRF) imaging is a promising technique for lymphatic mapping and sentinel lymph node (SLN) detection. The purpose of this technical feasibility pilot study was to evaluate the applicability of NIRF imaging with indocyanin green (ICG) for the detection of the SLN in cervical cancer.

Tumor-infiltrating Cytotoxic T Lymphocytes As Independent Prognostic Factor in Epithelial Ovarian Cancer with Wilms Tumor Protein 1 Overexpression

Journal of Immunotherapy (Hagerstown, Md. : 1997). Jul-Aug, 2011  |  Pubmed ID: 21654520

Immune response characterization at the primary tumor site enables the design of therapeutic vaccination strategies with higher efficacy in epithelial ovarian cancer (EOC). In this study, we related Wilms tumor protein 1 (WT1) overexpression, a well-established immunotherapeutic target, to clinicopathological characteristics, immunological parameters, and survival in primary EOC. WT1 overexpression was evaluated in primary EOC tissue of 270 patients by immunohistochemistry on tissue microarrays (TMAs). Clinicopathological characteristics, follow-up, and data on infiltration of CD8⁺ cytotoxic T lymphocytes (CTLs), FoxP3⁺ regulatory T lymphocytes (Tregs), major histocompatibility complex (MHC) class I, and II molecule expression, were derived from a previously published dataset. WT1 overexpression was defined as positive immunostaining for WT1. WT1 overexpression, present in 56.3% of EOC, was associated with infiltration of Tregs [odds ratio (OR), 2.7; 95% confidence interval (95% CI), 1.6-4.7; P<0.001] and up-regulation of MHC class II (OR, 2.2; 95% CI, 1.2-4.1; P=0.014). Advanced stage (OR, 4.0; 95% CI, 1.9-8.6; P<0.001) and serous histology (OR, 6.7; 95% CI, 3.2-13.6; P<0.001) were independent predictors of WT1 overexpressing EOC. High number of CTL was an independent prognostic factor for progression-free survival (hazard ratio, 0.5; 95% CI, 0.3-0.8; P=0.006) in WT1 overexpressing EOC. As WT1 overexpressing EOC is associated with CTL and Treg infiltration next to MHC class II up-regulation, future clinical trials should evaluate the combination of therapeutic WT1 vaccines with strategies depleting Tregs and/or up-regulating MHC class I, in an attempt to enhance clinical efficacy in EOC patients.

Training in Bowel and Upper Abdominal Surgery in Gynaecological Oncology: European Society of Gynecological Oncology (ESGO) Statement

International Journal of Gynecological Cancer : Official Journal of the International Gynecological Cancer Society. Oct, 2011  |  Pubmed ID: 21720255

Serum Tryptophan and Kynurenine Concentrations As Parameters for Indoleamine 2,3-dioxygenase Activity in Patients with Endometrial, Ovarian, and Vulvar Cancer

International Journal of Gynecological Cancer : Official Journal of the International Gynecological Cancer Society. Oct, 2011  |  Pubmed ID: 21720257

Indoleamine 2,3-dioxygenase (IDO) suppresses the function of T-lymphocytes and is involved in immune escape of cancers. Indoleamine 2,3-dioxygenase catalyzes the initial rate-limiting step in the degradation of the essential amino acid tryptophan. In this study, we investigated cancer-induced IDO activity in sera of endometrial, ovarian, and vulvar cancer patients.

Cell Surface Delivery of TRAIL Strongly Augments the Tumoricidal Activity of T Cells

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Sep, 2011  |  Pubmed ID: 21753155

Adoptive T-cell therapy generally fails to induce meaningful anticancer responses in patients with solid tumors. Here, we present a novel strategy designed to selectively enhance the tumoricidal activity of T cells by targeted delivery of TNF-related apoptosis-inducing ligand (TRAIL) to the T-cell surface.

Intraoperative Tumor-specific Fluorescence Imaging in Ovarian Cancer by Folate Receptor-α Targeting: First In-human Results

Nature Medicine. Oct, 2011  |  Pubmed ID: 21926976

The prognosis in advanced-stage ovarian cancer remains poor. Tumor-specific intraoperative fluorescence imaging may improve staging and debulking efforts in cytoreductive surgery and thereby improve prognosis. The overexpression of folate receptor-α (FR-α) in 90-95% of epithelial ovarian cancers prompted the investigation of intraoperative tumor-specific fluorescence imaging in ovarian cancer surgery using an FR-α-targeted fluorescent agent. In patients with ovarian cancer, intraoperative tumor-specific fluorescence imaging with an FR-α-targeted fluorescent agent showcased the potential applications in patients with ovarian cancer for improved intraoperative staging and more radical cytoreductive surgery.

Potentiation of a P53-SLP Vaccine by Cyclophosphamide in Ovarian Cancer: A Single-arm Phase II Study

International Journal of Cancer. Journal International Du Cancer. Dec, 2011  |  Pubmed ID: 22139992

The purpose of the current phase II single-arm clinical trial was to evaluate whether pretreatment with low-dose cyclophosphamide improves immunogenicity of a p53-synthetic long peptide (SLP) vaccine in patients with recurrent ovarian cancer. Patients with ovarian cancer with elevated serum levels of CA-125 after primary treatment were immunized four times with the p53-SLP vaccine. Each immunization was preceded by administration of 300 mg/m(2) intravenous cyclophosphamide as a means to affect regulatory T cells (Tregs). Vaccine-induced p53-specific interferon-gamma (IFN-γ)-producing T cells evaluated by IFN-γ ELISPOT were observed in 90% (9/10) and 87.5% (7/8) of evaluable patients after two and four immunizations, respectively. Proliferative p53-specific T cells, observed in 80.0% (8/10) and 62.5% (5/8) of patients, produced both T-helper 1 and T-helper-2 cytokines. Cyclophosphamide induced neither a quantitative reduction of Tregs determined by CD4(+) FoxP3(+) T cell levels nor a demonstrable qualitative difference in Treg function tested in vitro. Nonetheless, the number of vaccine-induced p53-specific IFN-γ-producing T cells was higher in our study compared to a study in which a similar patient group was treated with p53-SLP monotherapy (p ≤ 0.012). Furthermore, the strong reduction in the number of circulating p53-specific T cells observed previously after four immunizations was currently absent. Stable disease was observed in 20.0% (2/10) of patients, and the remainder of patients (80.0%) showed clinical, biochemical and/or radiographic evidence of progressive disease. The outcome of this phase II trial warrants new studies on the use of low-dose cyclophosphamide to potentiate the immunogenicity of the p53-SLP vaccine or other antitumor vaccines.

Anticancer Drugs Aimed at E6 and E7 Activity in HPV-positive Cervical Cancer

Current Cancer Drug Targets. Oct, 2011  |  Pubmed ID: 22165971

Standard treatment of locally advanced cervical cancer currently consists of concurrent chemoradiation, leading to a 5-year disease-free survival of 66-79%, indicating that there is still ample room for improvement. Characteristic of cervical cancer is the presence of high risk (HR) human papillomavirus (HPV) DNA in more than 99% of these tumors. When the HR HPV genome integrates in the host genome, oncogenic E6 and E7 proteins become constitutively expressed. These oncogenes are also active earlier in the infection cycle and hence are available as therapeutic targets at the preneoplastic stages as well. E7 plays an important role in the early stage of carcinogenesis by stimulating proliferation. HR HPV E6-induced proteasomal degradation of p53 hampers p53 functionality in cell cycle arrest and apoptosis. As p53 plays a key role in the intrinsic apoptotic pathway, current chemoradiation cannot optimally activate this pathway. In this review, we focus on targeted anticancer drugs to eliminate the consequences of HR HPV E6 and E7 activity. Strategies for direct and indirect targeting of HR HPV E6 and E7, including RNA interference, small molecules, proteasome inhibitors, and histone deacetylase inhibitors, are described. In addition, the extrinsic apoptotic pathway as possible alternative therapeutic target for apoptosis induction is reviewed. The rational for implementing recombinant human TRAIL and death receptor agonists and the latest developments on combining these drugs with standard treatment in preclinical settings as well as clinical trials are discussed.

ESGO Statement on the Role of CA-125 Measurement in Follow-up of Epithelial Ovarian Cancer

International Journal of Gynecological Cancer : Official Journal of the International Gynecological Cancer Society. Jan, 2012  |  Pubmed ID: 22193647

The Role of ATM and 53BP1 As Predictive Markers in Cervical Cancer

International Journal of Cancer. Journal International Du Cancer. Feb, 2012  |  Pubmed ID: 22323184

Treatment of advanced stage cervical cancers with (chemo)radiation causes cytotoxicity through induction of high levels of DNA damage. Tumour cells respond to DNA damage by activation of the 'DNA damage response' (DDR), which induces DNA repair and may counteract chemoradiation efficacy. Here, we investigated DDR components as potential therapeutic targets and verified the predictive and prognostic value of DDR activation in cervical cancer patients treated with (chemo)radiation. In a panel of cervical cancer cell lines, inactivation of ATM or its substrate 53BP1 clearly gave rise to cell cycle defects in response to irradiation. Concordantly, clonogenic survival analysis revealed that ATM inhibition, but not 53BP1 depletion, strongly radio-sensitised cervical cancer cells. In contrast, ATM inhibition did not radiosensitise non-transformed epithelial cells or non-transformed BJ fibroblasts. Interestingly, high levels of active ATM prior to irradiation were related with increased radioresistance. To test whether active ATM in tumours prior to treatment also resulted in resistance to therapy, immunohistochemistry was performed on tumour material of advanced stage cervical cancer patients (n=375), treated with (chemo)radiation. High levels of phosphorylated (p-)ATM (P=0.006, HR=1.817) were related to poor loco-regional disease-free survival. Furthermore, high p-ATM levels predicted shorter disease-specific survival (P=0.038, HR=1.418). Presence of p-53BP1 was associated with p-ATM (P=0.001, OR=2.206), but was not related to any clinicopathological features or survival. In conclusion, both our in vitro and patient-related findings indicate a protective role for ATM in response to (chemo)radiation in cervical cancer and point at ATM inhibition as a possible means to improve efficacy of (chemo)radiation. © 2012 Wiley-Liss, Inc.

HPV16 E6 RNA Interference Enhances Cisplatin and Death Receptor-mediated Apoptosis in Human Cervical Carcinoma Cells

Molecular Pharmacology. Feb, 2012  |  Pubmed ID: 22328720

In cervical cancer the p53 and pRb tumor suppressor pathways are disrupted by the human papilloma virus (HPV) E6 and E7 oncoproteins, since E6 targets p53 and E7 targets pRb for rapid proteasome-mediated degradation. We have investigated whether E6 suppression with small interfering (si)RNA restores p53 functionality and sensitizes the HPV16-positive cervical cancer cell line SiHa to apoptosis by cisplatin, irradiation, recombinant human tumor necrosis factor (TNF)-related apoptosis-inducing ligand (rhTRAIL) or agonistic anti-Fas antibody. E6 siRNA resulted in decreased E6 mRNA levels, enhanced p53 and p21 expression, demonstrating restoration of p53 functionality in SiHa, without inducing high levels of apoptosis (less than 10%). Cell surface expression of the proapoptotic death receptors DR4, DR5 and Fas was not affected by E6 suppression. E6 suppression conferred susceptibility to cisplatin-induced apoptosis but not to irradiation-, rhTRAIL- or anti-Fas antibody-induced apoptosis. Combining cisplatin with rhTRAIL or anti-Fas antibody induced even higher apoptosis levels in E6-suppressed cells. At the molecular level, cisplatin treatment resulted in elevated p53 levels, enhanced caspase-3 activation and reduced p21 levels in E6-suppressed cells. Cisplatin in combination with death receptor ligands enhanced caspase-8 and caspase-3 activation and reduced XIAP levels in these cells. We showed using siRNA that this enhanced apoptosis in E6-supressed cells was related to reduced XIAP levels and not due to reduced p21 levels. In conclusion, targeting E6 or XIAP in combination with cisplatin can efficiently potentiate rhTRAIL-induced apoptosis in HPV-positive cervical cancer cells.

The Effect of Chemotherapy on Expression of Folate Receptor-alpha in Ovarian Cancer

Cellular Oncology (Dordrecht). Feb, 2012  |  Pubmed ID: 21647742

Folate receptor alpha (FR-α) has been identified as a potential target in ovarian cancer for diagnostic and therapeutic purposes, based on its overexpression in serous epithelial ovarian carcinoma. The effect of chemotherapy on FR-α expression may be important in the applicability of FR-α directed agents in the case of residual tumor tissue. The objective of this study was to assess FR-α expression in ovarian carcinoma and to evaluate whether FR-α expression is altered by chemotherapy.