Preliminary Study On The Involvement Of Entpd5 In The Secondary Drug Resistance Of Cispatin In Epithelial Ovarian Cancer

Backgound and purposeEpithelial ovarian cancer is a malignant tumor that seriously threatens women's health.In the early stage of treatment,it is sensitive to cisplatin.However,with the deepening of treatment,the majority of patients have resistance to cisplatin and accompany tumor recurrence,resulting in a 5-year survival rate of patients with ovarian cancer is much lower than other Tumors are only 10-30%.Entpd5(ectonucleoside triphosphate dephosphohydrolase 5,also known as CD39L4)is a nucleotide hydrolase encoding the endoplasmic reticulum,which is mainly found in the endoplasmic reticulum and mediates intracellular nucleotide catabolism.Entpd5 has been found to mediate the survival of melanoma tumor cells,resist endoplasmic reticulum stress-mediated apoptosis,and promote tumor cell metastasis.In prostate cancer,the expression of Entpd5 is positively correlated with the degree of prostate cancer,and in the platinum-based chemotherapy resistance of prostate cancer,Entpd5 is associated with the stable state of Bcl-2 mediated by protein kinase Ca.Whether Entpd5 is involved in the regulation of drug resistance in platinum-based drugs for epithelial ovarian cancer has not been reported.This study was to investigate whether expression of Entpd5 in normal ovarian tissue,benign epithelial ovarian tumors,and epithelial ovarian cancer is different,and to further investigate whether Entpd5 is involved in the regulation of platinum drug resistance in epithelial ovarian cancer.Methods1.Expression of Entpd5 in benign and malignant epithelial ovarian tumor tissuesThe expression of Entpd5 in normal ovarian tissue,benign ovarian lesions and epithelial ovarian cancer was analyzed by immunohistochemistry by collecting specimens of normal ovarian tissue,benign ovarian lesions and epithelial ovarian cancer.2.The relationship between the expression level of Entpd5 and the resistance of epithelial ovarian cancer cells to cisplatin2.1 we analyzed the sensitivity of seven epithelial ovarian cancer cellsto Cisplatin by CCK8.Fluorescence quantitative PCR(q-PCR)was used to analyze the expression level of Entpd5 in different epithelial ovarian cancer cell lines.2.2 Ovcar8 and A1847 sensitive to cisplatin were selected to establish cisplatin secondary resistant cell lines Ovcar8R and A1847R.The drug sensitivity of CCK8 was detected in the parents and drug-resistant cells,and the expression level of Entpd5 was detected by Western blot.Results1.Expression of Entpd5 in benign and malignant epithelial ovarian tumor tissuesEntpd5 expression is weak or not expressed in normal ovarian tissue,and it is significantly increased in benign epithelial ovarian tumor and epithelial ovarian cancer tissues,and the expression of Entpd5 in epithelial ovarian cancer tissues and its benign epithelial ovarian tumor.The expression of tissues is significantly increased,so we believe that the expression of-Entpd5 is gradually increased with the deterioration of ovarian tissues,which is related to the degree of malignancy of ovarian tissues.2.The relationship between the expression level of Entpd5 and the resistance of epithelial ovarian cancer cells to cisplatin2.1 Cell level to verify the relationship between the sensitivity of different epithelial ovarian cancer cells to cisplatin and the expression level of Entpd5.Compared with the other five cell lines,the sensitivity of Ovcar8 and A1847 cells to cisplatin was significantly higher than that of other cells(P<0.01).Quantitative PCR(q-PCR)showed that different ovarian cancer cells expressed different degrees of Entpd5,and the expression of Entpd5 was increased according to A1847<Ovcar8<Ovcar3<Ovcar4<1GROV1<Ovcar5<SKOV3,and the difference was statistically significant.The results showed that the drug sensitivity of A1847 and Ovcar8 cells was significantly higher than that of the other five ovarian cancer cell lines,and the expression level of Entpd5 was significantly lower than that of other cells.2.2 Cisplatin-sensitive A1847 and Ovcar8 cell lines were selected to establish a corresponding drug-resistant cell lines model.CCK8 was used to detect drug sensitivity,and the expression level of Entpd5 in ovarian cancer cisplatin-resistant cells was confirmed by Western blot.The results showed that there were significant differences in the sensitivity of cisplatin between A1847 and Ovcar8 cells compared with A1847R and Ovcar8R cells(P<0.01).The expression of Entpd5 in drug-resistant cells was significantly higher than that in parental cisplatin-sensitive ovarian cancer cells,indicating that cisplatin-resistant in epithelial ovarian cancer cells are associated with increased expression of Entpd5.ConclusionEntpd5 has different expression levels in different degrees of ovarian lesions,it was significantly higher in epithelial ovarian cancer than in benign lesions in the diseased group.In epithelial ovarian cancer cells,cisplatin-resistant in epithelial ovarian cancer cells are associated with increased expression of Entpd5.In the cisplatin-sensitive parental cells,the expression level of Entpd5 was increased after the establishment of drug-resistant model,indicating that elevated expression of Entpd5 may promote the resistance of epithelial ovarian cancer to cisplatin.