LASP1 is an actin-binding protein associated with actin assembly dynamics in cancer cells. Here we report that LASP1 is overexpressed in pancreatic ductal adenocarcinoma (PDAC) where it promotes invasion and metastasis. We found that LASP1 overexpression in PDAC cells was mediated by HIF-1? through direct binding to a hypoxia response element in the LASP1 promoter. HIF-1? stimulated LASP1 expression in PDAC cells in vitro and mouse tumor xenografts in vivo. Clinically, LASP1 overexpression in PDAC patient specimens was associated significantly with lymph node metastasis and overall survival. Overall, our results defined LASP1 as a direct target gene for HIF-1? upregulation that is critical for metastatic progression of PDAC.
The aim of this study is to investigate the regulatory mechanism of leptin receptors (Ob-R) in pancreatic cancer. We found that the over-expression of hypoxia inducible factor (HIF-1)? and hypoxia up-regulated the expression of Ob-R in pancreatic cancer cells. When HIF-1? gene was silenced in vitro, the expression of Ob-R was significantly decreased. Xenograft mouse models showed that the inhibition of HIF-1? resulted in the concomitant decrease of Ob-R in vivo. In addition, HIF-1? expression was correlated with Ob-R in pancreatic cancer tissues by immunohistochemical staining. Clinical data showed that over-expression of HIF-1 was associated with pathological tumor node metastasis stage, lymph node metastasis and overall survival. HIF-1? directly bound to the hypoxia-responsive element (HRE) located in Ob-R gene promoter (-828/-832) and activated the transcription. Finally, we demonstrated that the silence of HIF-1? gene reversed the inhibitory effect of leptin/Ob-R in pancreatic cancer cells. Taken together, our results indicate that HIF-1? directly regulated Ob-R expression in pancreatic cancer, which might be a valuable therapeutic target for pancreatic cancer.
The aim of study was to discuss the correlation and regulatory mechanism of HIF-1 and miR-191 expression in pancreatic tumor. The association between the miR-191 and the clinicopathologic characteristics and the prognosis of pancreatic cancer was further explored. After hypoxic cultured for 6 and 12 h, qRT-PCR and Western blot were practiced to analyze the miR-191 and HIF-1 expression of MIA PaCa-2 and Aspac1 cells. We regulated the HIF-1 expression via plasmid and siRNA transfection to observe the alteration of HIF-1 and miR-191 expression. ChIP sequencing identified the binding sites of HIF-1 and miR-191. Dual luciferase assays were practiced to verify the binding sites. Immunohistochemical staining was practiced to analyze the expression of HIF-1, while qRT-PCR were done for investigating miR-191 in tumor tissues. Then, the association between the expression of them and the clinicopathologic characteristics and prognosis of pancreatic cancer were analyzed. After hypoxic cultured 12 h, the expression of HIF-1 protein, HIF-1mRNA and miR-191 of MIA PaCa-2 and AsPC-1 cells increased significantly (P?0.05). After HIF-1 overexpressing plasmid transfected to the MIA PaCa-2 and AsPC-1 cells for 48 h, the expression of HIF-1 protein, HIF-1mRNA, and miR-191 upregulated significantly (P?0.05). While after transfected the MIA PaCa-2 cells by HIF-1 siRNA, the significant decreasing of HIF-1 protein, HIF-1mRNA, and miR-191 expression were observed (P?0.05). ChIP sequencing showed the protein synthesis of HIF-1 increased in hypoxia situation. Only the HRE5 (-1,169 bp, ChIP4) were significantly brighter in hypoxia in comparing with normoxic cells. In dual luciferase assays, after pGL3-miR-191 and HIF-1 overexpressing plasmid co-transfect the MIAPaCa-2 cells for 48 h, its relative expression of bioluminescence was higher than those co-transfected by mutant miR-191 vectors and HIF-1 overexpressing plasmid or by pGL3-miR-191 and HIF-1 empty plasmid. The expression of miR-191 closely associated with the tumor size, pTNM stage, lymph node metastasis, and perineural invasion (P?0.05). Patients with higher expression of miR-191 were a risk factor for prognosis of pancreatic cancers. Expression of HIF-1 in pancreatic cancer cells increased under the condition of chronic hypoxia, which may bind to HRE2 in 5'flanking region of miR-191 and initiate transcription of miR-191. Expression of miR-191 was significantly higher in pancreatic tumor tissues. The expression of miR-191 closely associated with the tumor size, pTNM stage, lymph node metastasis and perineural invasion and poor prognosis of pancreatic cancer.
Stem cell factor (SCF), a ligand of c-kit, is a hematopoietic growth factor. Uncontrolled activity of SCF/c-kit signaling pathway contributes to the formation of a variety of human malignancies. In this study, we determined whether SCF expression could risk-stratify patients with hepatocellular carcinoma (HCC) after curative resection. HCC tissues from 160 patients were collected during curative resection and stained with SCF and CD34, a marker for microvessel density (MVD), using immunohistochemistry. Two statistical analyses were performed: an independent continuous and a multivariate categorical analysis, with test/validation set-defined cut points, and Kaplan-Meier estimated outcome measures of overall survival (OS) and relapse-free survival (RFS). We found that higher levels of SCF confer worse OS (continuous P = 0.014; and categorical P = 0.009), and RFS (continuous P = 0.002; categorical P = 0.003) of patients with HCC. SCF varies independently from MVD-CD34, tumor node metastasis, histologic grade, age and gender, and retains prognostic significance when analysed as a categorical variable in a multivariate analysis . We confirmed that MVD-CD34 is also an independent prognostic marker for patients with HCC. The levels of SCF and CD34 showed a positive and significant correlation (P < 0.0001) and double low expression confers superior OS (median = 48 months) and RFS (median = 24 months), whereas double high expression confers shortest RFS (median = 10.5 months) compared with single measurements. The prognostic values of SCF and CD34 were independently determined in this study and we propose that both of them are independent prognostic markers for HCC.
Because of the early onset of local invasion and distant metastasis, pancreatic ductal adenocarcinoma (PDAC) is the most lethal human malignant tumor, with a 5-year survival rate of less than 5%. In this study, we investigated the role of fascin, a prometastasis actin-bundling protein, in PDAC progression, invasion, and the molecular mechanisms underlying fascin overexpression in PDAC. Our data showed that the expression levels of fascin were higher in cancer tissues than in normal tissues, and fascin overexpression correlated with the PDAC differentiation and prognosis. Fascin overexpression promoted PDAC cell migration and invasion by elevating matrix metalloproteinase-2 (MMP-2) expression. Fascin regulated MMP-2 expression through protein kinase C and extracellular signal-regulated kinase. Importantly, our data showed that hypoxia induced fascin overexpression in PDAC cells by promoting the binding of hypoxia-inducible factor-1 (HIF-1) to a hypoxia response element on the fascin promoter and transactivating fascin mRNA transcription. Intriguingly, HIF-1? expression levels in PDAC patient specimens significantly correlated with fascin expression. Moreover, immunohistochemistry staining of consecutive sections demonstrated colocalization between HIF-1? and fascin in PDAC specimens, suggesting that hypoxia and HIF-1? were responsible for fascin overexpression in PDAC. When ectopically expressed, fascin was able to rescue PDAC cell invasion after HIF-1? knockdown. Our results demonstrated that fascin is a direct target gene of HIF-1. Our data suggested that the hypoxic tumor microenvironment in PDAC might promote invasion and metastasis by inducing fascin overexpression, and fascin might be targeted to block PDAC progression.
We investigated the incidence of primary pancreatic cancer with previous non-pancreatic cancer (PPC) and secondary metastatic cancer within the pancreas (SMC) to elucidate the differential diagnosis and treatment of these lesions.
The aim of this study was to identify the expression of prostate-specific membrane antigen (PSMA) and analyze the correlation between PSMA with clinical characteristics in patients with pancreatic cancer. The expression of PSMA protein and mRNA was detected by immunohistochemistry and real-time quantitative polymerase chain reaction in pancreatic cancer tissues, pancreatic intraepithelial neoplasia or normal pancreatic tissues, respectively. And clinical characteristics and prognosis of patients were investigated. PSMA was expressed in pancreatic cancer cells, both in protein and mRNA levels. Moreover, the PSMA levels were associated with the prognosis of patients with pancreatic ductal adenocarcinoma. The overall survival time of pancreatic cancer patients with high expression of PSMA was significantly shorter than that of the low ones. Moreover, the PSMA levels were correlated with clinicopathological features including the histological grade and pathological tumor-node-metastasis stage. PSMA is involved in the carcinogenesis of pancreatic cancer, and it might serve as a potential therapeutic target for pancreatic cancer.
Hypoxia-inducible factor-1? (HIF-1?) is a highly important transcription factor involved in cell metabolism. HIF-1? promotes glycolysis and inhibits of mitochondrial respiration in pancreatic ductal adenocarcinoma (PDAC). In response to tumor hypoxia, cyclophilin A (CypA) is over-expressed in various cancer types, and is associated with cell apoptosis, tumor invasion, metastasis, and chemoresistance in PDAC. In this study, we showed that both HIF-1? and CypA expression were significantly associated with lymph node metastasis and tumor stage. The expression of CypA was correlated with HIF-1?. Moreover, the mRNA and protein expression of CypA markedly decreased or increased following the suppression or over-expression of HIF-1? in vitro. Chromatin immunoprecipitation analysis showed that HIF-1? could directly bind to the hypoxia response element (HRE) in the CypA promoter regions and regulated CypA expression. Consistent with other studies, HIF-1? and CypA promoted PDAC cell proliferation and invasion, and suppressed apoptosis in vitro. Furthermore, we proved the combination effect of 2-methoxyestradiol and cyclosporin A both in vitro and in vivo. These results suggested that,CypA, a gene downstream of HIF-1?, could promote the development of PDAC. Thus, CypA might serve as a potential therapeutic target for PDAC.
This study aimed to investigate the interaction and regulatory mechanism of microRNA185 (miR185) and hypoxia-inducible factor-1 (HIF-1) in pancreatic cancer. The significance of miR185 on the clinicopathologic characteristics and prognosis was further explored. qRT-PCR and immunohistochemistry examined miR185 and HIF-1 expression of tumor tissues. Western blot analyzed HIF-1 protein expression. We regulated HIF-1 via transfection to observe the impact of HIF-1 on miR185 expression. ChIP sequencing and dual luciferase identified binding sites of HIF-1 and miR185. MiR185 expression was significantly higher in pancreatic tumors. MiR185 closely associated with tumor size, pTNM stage, lymph node, and perneural invasion. After hypoxic culture, both HIF-1 and miR185 expression of MiaPaCa2 and AsPc1 cells increased significantly. Up- or down-regulating HIF-1 expression via transfection leads to synchronous alteration of miR185. In ChIP sequencing, only the HRE2 (-938 bp) was significantly brighter under hypoxia among four hypoxia response elements (HREs) sequence of miR185 promoter. After pGL3-miR185 and HIF-1 over-expressing plasmids co-transfect the MiaPaCa2 cells, its relative expression of bioluminescence increased. MiR185 expression was significantly higher in tumor tissues and closely associated with the clinical features of pancreatic cancer. Expression of HIF-1 in pancreatic cancer cells increased in hypoxia. HIF-1 may bind to HRE2 of miR185 and initiate its transcription.
IL-35 is a novel inhibitory cytokine that is mainly produced by regulatory T-cells (Tregs) and is required for Treg-mediated immunosuppression. However, the plasma levels of IL-35 in patients with pancreatic ductal adenocarcinoma (PDAC) have never been investigated. In this study, we found that plasma IL-35 levels more significantly increased in PDAC patients than in normal controls (134.53±92.45pg/mL vs. 14.26±6.56pg/mL). IL-35 mRNA levels were positively correlated with plasma IL-35 levels (EBI3, R=0.925, p<0.01; p35, R=0.916, p<0.01). Furthermore, IL-35 expression levels were associated with lymph node metastasis (p=0.001) and late tumor stage (p=0.002). For the resected patients, high IL-35 expression levels were associated with large tumor size (p<0.01), higher TNM classification T staging (p<0.05), and late tumor stage (p<0.05). In conclusion, circulating IL-35 in PDAC patients significantly increased, suggesting that regulating the expression of IL-35 may provide a new possible target for the treatment of PDAC patients, especially for the resectable ones.
One of the hallmarks of cancer is revised glucose metabolism that promotes cell survival and proliferation. In pancreatic cancer, the regulatory mechanism of glucose metabolism remains to be elucidated. In this study, we found that CX3CR1 is expressed in pancreatic cancer cells lines. Exogenous or transfected CX3CL1 increased glucose uptake and lactate secretion. CX3CL1 stimulated HIF-1 expression through PI3K/Akt and MAPK pathways. Furthermore, knockdown of HIF-1 blocked CX3CL1-modified glucose metabolism in pancreatic adenocarcinoma cells. In conclusion, the CX3CL1/CX3CR1 reprograms glucose metabolism through HIF-1 pathway in pancreatic cancer cells.
Parkin, an E3 ubiquitin ligase well known for its role in the pathogenesis of juvenile Parkinson disease, has been considered as a candidate tumor suppressor in certain types of cancer. It remains unknown whether parkin is involved in the development of pancreatic cancer, the fourth leading cause of cancer-related deaths worldwide. Herein, we demonstrate the downregulation and copy number loss of the parkin gene in human pancreatic cancer specimens. The expression of parkin negatively correlates with clinicopathological parameters indicating the malignancy of pancreatic cancer. In addition, knockdown of parkin expression promotes the proliferation and tumorigenic properties of pancreatic cancer cells both in vitro and in mice. We further find that parkin deficiency increases the proportion of cells with spindle multipolarity and multinucleation. Parkin-depleted cells also show a significant increase in spindle misorientation. These findings indicate crucial involvement of parkin deficiency in the pathogenesis of pancreatic cancer.
The transcription factor hypoxia-inducible factor-1 (HIF-1) has ? and ? subunits. Recent studies have shown that the HIF-1? gene may have C1772T and G1790A single nucleotide polymorphisms (SNPs). These SNPs may increase the stability and activity of HIF-1?. In the present study, we looked for these SNPs by genotyping circulating mononuclear cells from 263 patients with pancreatic ductal adenocarcinoma (PDAC), using 271 healthy volunteers as controls. As a result, both SNPs were more frequent in PDAC patients than in healthy volunteers (C1772T: 21 vs. 11%, p < 0.01; G1790A: 25 vs. 8%, p < 0.01). Further, both SNPs were associated with higher risks for PDAC (C1772T: OR=2.156, 95% CI: 1.324-3.511, p < 0.05; G1790A: OR=3.716, 95% CI: 2.213-6.238, p < 0.01). We also stained HIF-1? by immunohistochemistry in 68 PDAC tumors to examine their HIF-1? expression levels. To this end, we designed a semi-quantitative method that was based on the staining intensity and frequency of HIF-1?-positive cells. As a result, the G1790A SNP, but not C1772T SNP, was associated with an increased HIF-1? expression. We also related genotyping data to patients survival times, serum CA19-9, and tumors volumes, grades, stages and lymph-node metastasis. The C1772T SNP was not associated with any of these parameters. In contrast, the G1790A SNP was associated with increases in serum CA19-9 and in tumor volumes. In conclusion, the C1772T and G1790A SNPs in the HIF-1? gene increase the susceptibility to pancreatic cancer. In addition, the G1790A SNP is associated with increases in tumor-produced HIF-1? and in the progression of the cancer.
Tumor rupture is considered a R2 resection and is not uncommonly encountered when attempting a tumor-free resection, especially in high-risk soft tissue sarcomas. The predictive value of tumor rupture for metastasis development and its impact on metastasis-free interval needs to be investigated and further taken into consideration in preoperative planning.
The incidence of pancreatic cancer has increased in China in the last decade, though efforts have been made in early detection and multimodality treatment. The aim of this study is to describe the decade-based development in early diagnosis and treatment modalities, as well as outcome for patients with pancreatic ductal adenocarcinoma (PDAC) in a high-volume facility.
Pancreatic cancer is a highly aggressive disease with a grim prognosis, due to its late diagnosis, propensity to rapidly metastasize, and resistance to therapy. The molecular events underlying this remain poorly defined. Here we report the overexpression and gene copy number gain of the microtubule-dependent motor protein Eg5 in human pancreatic cancer samples. We also show that Eg5 expression correlates with clinicopathological parameters of pancreatic cancer and promotes anchorage-independent cell growth and tumour formation in mice. In addition, Eg5 is up-regulated in pancreatic cancer cell lines and enhances cell proliferation in an ATPase activity-dependent manner. Our data further reveal that Eg5 overexpression causes the formation of multipolar spindles and multinucleation and induces the accumulation of polyploid cells. These findings demonstrate a role for Eg5 in pancreatic tumourigenesis and indicate a potential for targeting Eg5 in pancreatic cancer treatment.
The aim was to analyze the mechanism of leptin-induced activity of telomerase in MCF-7 breast cancer cells. We found that leptin activated telomerase in a dose-dependent manner; leptin upregulated the expression of Human Telomerase Reverse Transcriptase (hTERT) at mRNA and protein levels; blockade of signal transducer and activator of transcription 3 (STAT3) phosphorylation significantly counteracted leptin-induced hTERT transcription and protein expression; chromatin immunoprecipitation analysis showed that leptin enhanced the binding of STAT3 to the hTERT promoter. This study uncovers a new mechanism of the proliferative effect of leptin on breast cancer cells and provides a new explanation of obesity-related breast cancer.
CX3CR1 is an important chemokine receptor and regulates the chemotactic migration of pancreatic ductal adenocarcinoma (PDAC) cells. Up to now, its regulatory mechanism remains largely undefined. Here, we report that hypoxia upregulates the expression of CX3CR1 in pancreatic cancer cells. When hypoxia-inducible factor (HIF)-1? expression was knocked down in vitro and in vivo, the expression of CX3CR1 was significantly decreased. Chromatin immunoprecipitation assay demonstrated that HIF-1? bound to the hypoxia-response element (HRE; 5-A/GCGTG-3) of CX3CR1 promoter under normoxia, and this binding was significantly enhanced under hypoxia. Overexpression of HIF-1? significantly upregulated the expression of luciferase reporter gene under the control of the CX3CR1 promoter in pancreatic cancer cells. Importantly, we demonstrated that HIF-1? may regulate cancer cell migration through CX3CR1. The HIF-1?/CX3CR1 pathway might represent a valuable therapeutic target to prevent invasion and distant metastasis in PDAC.
To investigate the efficacy and safety of regional intra-arterial chemotherapy (RIAC) versus systemic chemotherapy for stage III/IV pancreatic cancer.
Related JoVE Video
Journal of Visualized Experiments
What is Visualize?
JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.
How does it work?
We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.
Video X seems to be unrelated to Abstract Y...
In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.