method-for-obtaining-primary-ovarian-cancer-cells-from-solid-specimens

Method for Obtaining Primary Ovarian Cancer Cells From Solid Specimens

Reliable tools for investigating ovarian cancer initiation and progression are urgently needed. While the use of ovarian cancer cell lines remains a ...

University of Minnesota

A 31-year-old female developed multiple episodes of grand mal seizures after combination chemotherapy with cisplatin, vinblastine and bleomycin for germ cell ovarian cancer stage Ic. The clinicoradiologic features in this patient were consistent with posterior leukoencephalopathy, which is a rare complication of chemotherapy. Seizures were controlled by the anticonvulsive agent Dilantin (Pfizer, Khet Klongtoey, Bangkok) and she returned home without any permanent neurologic deficits. Follow-up magnetic resonance imaging 2 weeks later showed complete resolution of the abnormalities. This syndrome has been previously reported following cisplatin-based chemotherapy. Physicians should remain alert to the potential hazards of chemotherapy to the central nervous system. Risks and benefits should be seriously considered before starting treatment.

Posterior leukoencephalopathy following cisplatin, bleomycin and vinblastine therapy for germ cell tumor of the ovary.

Pregnancy in the broad ligament is a rare form of ectopic pregnancy, and one type of abdominal pregnancy. The diagnosis is seldom established before surgery. A 38-year-old, 11-week pregnant woman, gravida 3, para 2, presented with vaginal bleeding. She had undergone two cesarean sections 10 and 6 years earlier. Pregnancy in the right broad ligament was diagnosed from clinical and transvaginal ultrasonographic findings. Emergency laparotomy and excision of a pregnancy in the right broad ligament and right salpingectomy were performed. She was well at discharge and at the 6-week follow up. We suggest the use of clinical and ultrasonographic findings for the suspicion of pregnancy in the broad ligament.

Pregnancy in the broad ligament.

Benign cystic teratomas are one of the most common benign ovarian neoplasms. Although its rupture is rare, once occurred it can cause complications such as granulomatous peritonitis, mimicking metastatic ovarian malignancy.

Ovarian teratoma with diffused peritoneal reactions mimicking advanced ovarian malignancy.

To investigate the prevalence of abnormal Papanicolaou smear in pregnant patients who attend the Antenatal Care Clinic at Thammasat University Hospital.

Prevalence and management of abnormal pap smear in antenatal care clinic at Thammasat University Hospital.

A total of 74 patients with apparent early stage epithelial ovarian cancer who underwent exploratory laparotomy at King Chulalongkorn Memorial Hospital or other hospitals and were referred for further treatment, were evaluated. Formalin fixed paraffin-embedded ovarian tissue specimens were collected and immuno-stained with HER-2/neu antibodies for comparison with clinicopathologic data after median follow up of 46 months (range 3 - 83 months). The prevalence of HER-2/neu overexpression in these patients was 10.2%. No significant correlation between HER-2/neu overexpression and clinicopathological parameters (stage, ascites, capsular rupture, capsular adherence, histological subtype and histological grade) was found. Disease free survival and overall survival did not statistically differ between those with lesions positive or negative for HER-2/neu overexpression.

Lack of prognostic significance of HER-2/neu in early epithelial ovarian cancer.

Ovarian cancer: risks.

Ovarian cancer is the deadliest gynecologic cancer. Unlike many cancers such as breast, cervical and colon cancers, there is no easily clinically identifiable pre-malignant phase of this malignancy making early identification difficult. Similarly, unlike lung, head and neck, and skin cancers, there is not easily identifiable risk factor making prevention short of oophorectomy difficult. Even so, theories as to the causative factors of ovarian cancer continue to evolve making our understanding of the genesis of ovarian cancer more clear. Genetics, parity, environment, hormonal factors, and inflammation all play an important and pivotal role in the development of ovarian cancer. The most current understanding of these elements and their respective contribution to the development of this cancer are presented in this chapter.

Current understanding of risk factors for ovarian cancer.

Solitary fibrous tumor is a rare mesenchymal tumor reported initially in the pleura but that is now reported in widely ranging anatomic sites with a variable clinical course. Solitary fibrous tumor arising from the female genital tract is extremely rare and the management of this condition is controversial.

Solitary fibrous tumors arising from the female pelvis.

Cervical cancer cells exhibit an increased requirement for ubiquitin-dependent protein degradation associated with an elevated metabolic turnover rate, and for specific signaling pathways, notably HPV E6-targeted degradation of p53 and PDZ proteins. Natural compounds with antioxidant properties including flavonoids and triterpenoids hold promise as anticancer agents by interfering with ubiquitin-dependent protein degradation. An increasing body of evidence indicates that their α-β unsaturated carbonyl system is the molecular determinant for inhibition of ubiquitin-mediated protein degradation up-stream of the catalytic sites of the 20S proteasome. Herein we report the identification and characterization of a new class of chalcone-based, potent and cell permeable chemical inhibitors of ubiquitin-dependent protein degradation, and a lead compound RAMB1. RAMB1 inhibits ubiquitin-dependent protein degradation without compromising the catalytic activities of the 20S proteasome, a mechanism distinct from that of Bortezomib. Treatment of cervical cancer cells with RAMB1 triggers unfolded protein responses, including aggresome formation and Hsp90 stabilization, and increases p53 steady state levels. RAMB1 treatment results in activation of lysosomal-dependent degradation pathways as a mechanism to compensate for increasing levels of poly-ubiquitin enriched toxic aggregates. Importantly, RAMB1 synergistically triggers cell death of cervical cancer cells when combined with the lysosome inhibitor Chloroquine.

Stressing the ubiquitin-proteasome system without 20S proteolytic inhibition selectively kills cervical cancer cells.

Ovarian cancer is the deadliest of the gynecological diseases and the fifth cause of cancer death among American women. This is mainly due to the lack of prognostic tools capable of detecting early stages of ovarian cancer and to the high rate of resistance to the current chemotherapeutic regimens. In this scenario the overall 5-year survival rate for ovarian cancer patients diagnosed at late stage is less than 25%. Abnormalities associated with the malignant phenotype and the mechanisms of tumor progression are not clearly understood. In vitro studies are necessary, yet have been hampered due to the limitations accompanied with the use of ovarian cancer cell lines and the heterogeneity of the ovarian cancer cell population derived from ascites fluids. In this study we present a simple, rapid and reproducible method for the isolation and characterization of ovarian cancer cells from solid tumor tissue and show that enzymatic digestion for 30 minutes with dispase II results in the most effective recovery of viable epithelial ovarian cancer (EOC) cells. The resulting cancer (EOC) cell preparations demonstrate a significant yield, high levels of viability and are fibroblast-free. They grow for up to six passages and retain the capacity of forming spheroids-like structures in agarose. In addition, they can be genetically manipulated and used for drug screening, thus rendering them highly suitable for downstream applications. Notably, isolation of ovarian cancer cells from solid specimens using this method has the advantage of allowing for isolation of cancer cells from early stages of ovarian cancer as well as obtaining cells from defined either primary and/or metastatic ovarian cancer sites. Thus, these cells are highly suitable for investigations aimed at understanding ovarian cancer.

Establishment, characterization and downstream application of primary ovarian cancer cells derived from solid tumors.

We aimed to evaluate the feasibility and tolerability of hyperthermic intraperitoneal carboplatin (HIPEC-carboplatin) following secondary cytoreduction for recurrent, platinum-sensitive ovarian cancer.

Hyperthermic intraperitoneal chemotherapy with carboplatin for optimally-cytoreduced, recurrent, platinum-sensitive ovarian carcinoma: a pilot study.

To assess the feasibility of using fat-fraction imaging for measuring marrow composition changes over large regions in patients undergoing cancer therapy.

Water-fat MRI for assessing changes in bone marrow composition due to radiation and chemotherapy in gynecologic cancer patients.

Anti-estrogen therapy appears to have efficacy in a subset of ovarian cancers, as demonstrated in multiple phase II studies. Identifying sensitive patients early in treatment may allow for targeted, low-toxicity primary therapy or prevention of recurrence. We have previously demonstrated that the likelihood of response to letrozole could be improved by patient selection based on estrogen-pathway marker expression. We sought to identify ovarian cancer biomarkers that might indicate sensitivity to fulvestrant, an estrogen receptor antagonist.

Article	Year
Posterior leukoencephalopathy following cisplatin, bleomycin and vinblastine therapy for germ cell tumor of the ovary. The journal of obstetrics and gynaecology research\| PubMed ID: 12078977	2002
Pregnancy in the broad ligament. Archives of gynecology and obstetrics\| PubMed ID: 12942257	2003
Ovarian teratoma with diffused peritoneal reactions mimicking advanced ovarian malignancy. Archives of gynecology and obstetrics\| PubMed ID: 15526210	2004
Prevalence and management of abnormal pap smear in antenatal care clinic at Thammasat University Hospital. Journal of the Medical Association of Thailand = Chotmaihet thangphaet\| PubMed ID: 15962660	2005
Lack of prognostic significance of HER-2/neu in early epithelial ovarian cancer. Asian Pacific journal of cancer prevention : APJCP\| PubMed ID: 18260719	2007
Ovarian cancer: risks. Hawaii medical journal\| PubMed ID: 19385377	2009
Current understanding of risk factors for ovarian cancer. Current treatment options in oncology\| PubMed ID: 19603272	2009
Solitary fibrous tumors arising from the female pelvis. Obstetrics and gynecology\| PubMed ID: 21768857	2011
Stressing the ubiquitin-proteasome system without 20S proteolytic inhibition selectively kills cervical cancer cells. PloS one\| PubMed ID: 21909374	2011
Establishment, characterization and downstream application of primary ovarian cancer cells derived from solid tumors. PloS one\| PubMed ID: 23226302	2012
Hyperthermic intraperitoneal chemotherapy with carboplatin for optimally-cytoreduced, recurrent, platinum-sensitive ovarian carcinoma: a pilot study. Gynecologic oncology\| PubMed ID: 23352917	2013
Water-fat MRI for assessing changes in bone marrow composition due to radiation and chemotherapy in gynecologic cancer patients. Journal of magnetic resonance imaging : JMRI\| PubMed ID: 23450703	2013
Predicting response to the anti-estrogen fulvestrant in recurrent ovarian cancer. Gynecologic oncology\| PubMed ID: 23911795	2013

Thanasak Sueblinvong

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