The Role Of Heat Shock Factor1in Invasion And Metastasis Of Endometrial Cancer

BackgroudEndometrial cancer is a group of endometrial epithelial malignant tumors,endometrial adenocarcinoma is the most common lesions. Endometrial cancer is theseventh most common malignant disease, and one of the three common malignant tumorsin female genital tract, accounting for7%of the female malignant tumor and about20%to30%in gynecological oncology, the incidence is on rise in recent years and variesamong regions. Substantial decreases in the incidence and mortality of endometrial cancerare unlikely in the next few years, because early detection and treatment strategies havenot had a great influence on mortality. Therefore, there is an urgent requirement for newbiomarkers for endometrial cancer, so as to develop better diagnosis and treatment ofendometrial carcinoma.Heat shock factors (HSF) contain four subtypes, namely HSF1, HSF2, HSF3, HSF4.As the most important transcription factor of Heat shock response, HSF1containing529amino acids, constituted by DNA binding domain (DBD), trimerization domain(TD),regulatory domain(RD), and transcriptional activation domain(TAD), including two extremely conservative nuclear area–TD and DBD. The common assumption is that theseeffects are mediated through regulation of heat-shock protein (HSP) expression. However,the transcriptional network that HSF1coordinates directly in malignancy and itsrelationship to the heat-shock response have never been defined.In recent years, the studyfound that HSF1is not only by regulating the HSPs genes also by cancer-causing genesindividually or jointly participate in the occurrence of malignant tumor. Researchesfounded that HSF1is highly expressed in a variety of malignant tumors, such as coloncancer, breast cancer, prostate cancer, liver cancer, lung cancer, etc. It is closely related tothe development and invasion and metastasis of tumors. High expression of HSF1associated with estrogen also related to the prognosis of breast cancer patients. So weconsider HSF1may be a potential target of endometrial carcinoma and can develop betterdiagnosis and treatment of endometrial cancer.There is few study about HSF1expression and function in endometrial cancer and weplan to explore HSF1expression in endometrial carcinoma and the related mechanism.This is the first report that has examined the expression HSF1in endometrial cancertissues and cell lines. We also investigated the correlations between HSF1expression andvarious clinicpathologic parameters and ER. Furthermore, we determined the functionalroles of HSF1in endometrial cancer cell lines. The results presented would help toevaluate the suitability of HSF1as a new biomarker.Objective1. To study the relationship between HSF1protein expression and clinical pathologyof endometrial cancer.2. To study HSF1expression in endometrial cancer cell lines.3.To study the potential correlation between HSF1and invasion and metastasis andapoptosis in endometrial cancer cells.Methods1. Immunohistochemistry technique were used to explore the relationship betweenHSF1expression and clinical pathology of endometrial cancer.2. qRT-PCR and Western-blot were used to explore HSF1and ER expression in endometrial cancer cell lines.3.To evaluate possible contributions of HSF1to cell migration and invasion andapoptosis, HSF1-siRNA were transfected into Ishikawa cells. The application of qRT-PCR and western blot to screen effective small interference RNA.4.qRT-PCR ere used to explore the mRNA of HSp90,MMP2and MMP9andWestern-blot were used to explore the protein of E-cadherin and β-catenin aftertransfection of HSF1-siRNA.5.The cell cycle and apoptosis of endometrial cancer cells were analyzed by Flowcytometry instrumentResults1.The relationship between HSF1expression and clinicopathological parameters of inthe endometria cancer tissue:a)We described the relationship between HSF1expression and clinicopathologicalprameters of135cases of endometrial cancer with immunohistochemical analysis, andwas localised in the nucleus of tumor cellsb)High HSF1expression was associated with a significantly Histological grade,Muscular invasion,Lymphnoed metastisi and ER+/-(P<0.05), but not with age, FIGOstage (P>0.05).2.QRT-PCR and western blot analysis showed that High HSF1expression inIshikawa, HEC–1B, RL95-2and mesenchymal cells (normal endometrial cancer cells) inturns (P <0.05); low ER expression in Ishikawa cell.3.The application of qRT-PCR and western blot to screen effective small interferenceRNA. The expression of HSF1protein and mRNA was decreased in HSF1-siRNA-transfected cells compared with control siRNA-transfected cells and untransfected cells(P<0.05).a)After transfection of cells, qRT-PCR was performed to analyze the mRNAexpression levels of HSP90,MMP2and MMP9. Western blot analysis the expression ofE-cadherin protein was increased,and β-catenin protein was declined in HSF1-siRNAtransfected cells compared with negative control cells and blank cells (P<0.05). b)The results showed a significant reduction in motility of HSF1-siRNA-transfectedIshikawa cells compared with the control groups (P<0.05).c) Down-rugulated of HSF1can inhibit invasive ability in HSF1-siRNA transfectedcells compared with negative control cells and blank cells (P<0.05).d)Down-rugulated of HSF1can increase the percentage of cells in G1phase andreduce the percentage of cells in S phase,also increase the percentage of cells of apoptosisin HSF1-siRNA transfected cells compared with negative control cells and blank cells (P<0.05).Conclusions1. This study is the first to examine the expression of HSF1in endometrial cancertissues.Our results indicated that High HSF1expression was associated with asignificantly Histological grade, Muscular invasion,Lymphnoed metastisi and ER+/-(P<0.05), but not with age, FIGO stage (P>0.05). This suggests that Basigin-2plays animportant role in endometrial cancer.2. There are high expression of HSF1in Ishikawa cells and low expression of ER.3.qRT-PCR and western blot showed HSF1may be related to E-cadherin, beta-catenin, MMP2, MMP9, HSP90interaction involved in tumor invasion and metastasis.4.The result of experiment showeHSF1can promote ovarian cancer cell migration,invasion, and play a more important function in the progression of endometrial cancer.