A Research Of The Function And Mechanism Of Potential Genes ET-1 And Ambral In Prostate Cancer

Objective:The incidence of prostate cancer is ranked second in male cancers,and fatality rate is ranked sixth.In recent years,a number of biomolecular markers related to the diagnosis and treatment of prostate cancer have been screened through extensive sequencing work.The full study and identification of these molecular markers will promote their accurate application in the diagnosis and treatment of cancer.It has been reported that the vasoconstrictive peptide Endothelin-1(ET-1)produced by the prostate epithelium may play an important role in the progression of prostate cancer.A novel ATG gene,Beclinl regulating Activating Molecule in Beclin1-Regulated Autophagy1(Ambra1),also regulates cisplatin sensitivity in PCa cells.This study focuses on the role of ET-1 in the progression of prostate cancer and the sensitivity of Ambral in prostate cancer through mediating autophagy.Methods:Western blotting was used to confirm the expression of ET-1 in prostate cancer cell lines(LNCap cell line,PC3 cell line and DU 145 cell line).In addition,we used siRNA to knock down the expression of ET-1 gene in human prostate cancer cell lines,ET-1 gene in human prostate cancer cell line cell proliferation,apoptosis,cell cycle and cell invasion and migration.Western blot,apoptosis and caspase 3 activity assay,fluorescein reporter system,colony formation assay and other methods to determine whether the Ambral promotes cisplatin-induced DU 145 cell autophagy,whether the improvement on Ambral improve cisplatin treatment DU 145 cell growth,whether Ambral in human PCa DU 145 cells could Inhibit Cisplatin-induced Apoptosis.Result:Our study revealed that ET-1 expression was cell-specific in tumors,and ET-1 expression was highest in PC3 cell lines,but expression was significantly lower(p<0.05).ET-1 knockdown causes human prostate cancer PC3 cell line cells to arrest mainly in G1 phase,the number of cells passing the G1/G2 checkpoint is significantly reduced compared to the control group(p<0.05);ET-1 knockdown can cause human prostate.The proportion of apoptosis in PC3 cell line was significantly higher than that in the control group(p<0.05).Cell invasion assays demonstrated that ET-1 knockdown caused a significant decrease in cell migration of human prostate cancer PC3 cell line and was more aggressive than the control group.The number of cells was significantly decreased(p<0.05)knockdown of ET-1 caused a decrease in the expression of RhoA protein associated with cell invasion and migration,and increased expression of a-catenin,E-cadherin,and RhoC proteins,suggesting that ET-1 gene pairs cells with the role of invasion and migration.In addition,we transfected the Ambral plasmid in human PCa DU 145 cells and found that the apoptosis rate induced by cisplatin was significantly aggravated to more than 20%.Ambral negatively regulates cisplatin-induced apoptosis in DU 145 cells.Ambral promotes autophagy in cisplatin-treated DU 145 cells.Through colony formation assay,we explored the regulation of cisplatin-treated DU145 cells by Ambral and found that Ambral can improve the growth of DU 145 cells treated with cisplatin.The conclusion:Our research shows that ET-1 gene plays a role in cell proliferation,cell apoptosis,cell cycle,cell invasion and migration in human prostate cancer cell lines,and a deeper analysis of these functions will promote our Knowledge of the Potential Role of the ET-1 Gene in the Diagnosis and Treatment of Human Prostate Cancer Cell Lines.In addition,we found that Ambral negatively regulates cisplatin-induced apoptosis and cisplatin-mediated growth reduction in DU 145 cells,and this negative regulation is associated with Ambral-mediated promotion of autophagy but not with the p62 signaling pathway.This means that Ambral-mediated autophagy may be an important mechanism leading to decreased sensitivity of PCa cells.