The Effect Of ER Stress And Autophagy In Apoptosis Induced By Cisplatin In NRK-52E

Cisplatin has significant anticancer effects for different types of tumors.However,its side effects,especially kidney toxicity,limit its clinical application.After cisplatin enters the kidney,it can induce apoptosis of renal tubular epithelial cells(NRK-52E)and produce renal toxicity.In order to study the mechanism of cisplatin induced apoptosis in NRK-52 E cells,we design the following ideas:Objective:In this study,cisplatin nephrotoxicity was induced by model that cisplatin induced NRK-52 E cells injury in vitro,and the mechanism of cisplatin induced apoptosis in renal tubular epithelial cells was studied.To observe the apoptosis of renal tubular epithelial cells after endoplasmic reticulum stress inhibitor sodium taurocholodeoxycholate(TUDCA)and autophagy inhibitor chloroquine(CQ),so as to lay a theoretical foundation for their therapeutic targets.Methods:1.The effect of cisplatin on the cell viability of NRK-52 E cells by MTT method;2.Annexin V-FITC/PI double staining assay detect the apoptosis rate of NRK-52 E cells;3.Western blot was uesd to observe the expression of cleaved caspase-3,GRP78,CHOP,and LC3-II;4.Cell proliferation was detected by RTCA;5.Hoechst staining assay observe nuclear morphology.Results:MTT assay showed that the effect of cisplatin on cell viability was dose-dependent;flow cytometry showed that cisplatin and cisplatin combined with TUDCA and CQ can increase the apoptosis rate of NRK-52 E cells;Western blot showed that the expression of cleaved caspase-3,GRP78,CHOP,LC3-II in cisplatin group was higher than the control group;RTCA showed that comparing with the growth curve of the control group,cisplatin group,TUDCA+cisplatin group,andCQ+cisplatin group decreased significantly,and TUDCA+cisplatin group and CQ+cisplatin group decreased more significantly;Hoechst staining showed that compared with the control group,the number of apoptotic cell of the cisplatin group,TUDCA+cisplatin group and CQ+cisplatin group increased.Conclusions:1.Cisplatin induced renal tubular epithelial cell injury caused endoplasmic reticulum stress and autophagy.2.Inhibition of endoplasmic reticulum stress and autophagy can significantly increase cisplatin induced damage to renal tubular epithelial cells,and induce apoptosis.