miRNA-200c逆转胃癌SGC7901/DDP细胞对顺铂的耐药性及其相关机制

目的:探讨微RNA-200c (microRNA-200c,miRNA-200c)在逆转胃癌SGC7901/DDP细胞对顺铂(cisplatin,DDP)耐药中的作用及其可能的作用机制.方法:应用实时荧光定量PCR (real-time fluorescence quantitative-PCR,RFQ-PCR)法检测对DDP耐药的胃癌细胞SGC7901/DDP及其亲本细胞SGC7901中miRNA-200c的表达差异,同时检测SGC7901/DDP细胞转染miRNA-200c前体片段后miRNA-200c表达的变化;应用MTT法检测转染后SGC7901/DDP细胞对DDP药物敏感度的改变;Western印迹法检测转染后细胞bax、10号染色体同源丢失性磷酸酶与张力蛋白(phosphatase and tensin homology deleted on chromosome ten,PTEN)和E-钙黏蛋白(E-cadherin,E-cad)的表达变化.结果:SGC7901/DDP细胞中miRNA-200c的相对表达量显著低于DDP敏感细胞株SGC7901中的表达量 (P<0.05),SGC7901/DDP细胞转染miRNA-200c前体片段24 h后细胞中miRNA-200c的表达水平比转染阴性对照片段的细胞明显提高(P<0.05).DDP对转染miRNA-200c前体片段细胞的半数抑制浓度(half inhibition concentration,IC50)显著低于转染阴性对照片段的细胞(P<0.05).转染miRNA-200c前体片段后,SGC7901/DDP细胞的bax、PTEN和E-cad的蛋白表达水平明显高于转染阴性对照片段的细胞 (P<0.05).结论:miRNA-200c可能通过上调E-cad、PTEN和bax蛋白表达而逆转SGC7901/DDP细胞对DDP的耐药.

Reversion of the resistance of gastric cancer SGC7901/DDP cells by miRNA-200c and its related mechanism

Objective:To explore the effect of microRNA-200c(miRNA-200c)in reversing cisplatin(DDP)resistance of gastric cancer SGC7901/DDP cell line and elucidate its related mechanisms.Methods:The real-time fluorescence quantitative PCR(RFQ-PCR)was used to detect miRNA-200c expression in SGC7901 cells,DDP-resistant SGC7901/DDP cells,and miRNA-200c transfected SGC7901/DDP cells.The difference in miRNA-200c expression was compared between the groups.Drug sensitivity to DDP after transfection was tested using MTT assay....