microRNA let-7e逆转人卵巢癌细胞顺铂耐药及其作用机制  

Effect and mechanism of the reverse of the cisplatin resistance in human ovarian cancer cells by microRNA let-7e

作　　者：杨纯[1] 蔡晶[1] 胡沙[1] 王齐月 杨强[1] 王泽华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院,武汉430022

出　　处：《现代妇产科进展》2014年第11期868-871,共4页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金青年基金项目(No:81101961)

摘　　要：目的:研究人类microRNA let-7e(hsa-let-7e)在人卵巢癌顺铂耐药细胞中的表达及其作用机制。方法:采用实时荧光定量PCR法检测let-7e在卵巢癌亲本细胞株A2780及耐顺铂卵巢癌细胞A2780/CP中的表达;采用实时荧光定量PCR法和Western blot法分别检测A2780细胞和A2780/CP细胞中果蝇zeste基因增强子同源物2(EZH2)mRNA和蛋白的表达。将let-7e模拟物(mimics)及阴性对照(negative control,NC)转染A2780/CP细胞,采用实时荧光定量PCR和Western blot法分别检测EZH2 mRNA和蛋白的表达,MTT法检测细胞的半数抑制浓度(IC50),台盼蓝染色实验检测经顺铂作用不同时间后的细胞存活率。结果:与A2780细胞比较A2780/CP细胞中let-7e呈低表达、EZH2mRNA和蛋白相对表达水平显著增高,差异均有统计学意义(P<0.05)。A2780/CP细胞转染let-7e mimics后,EZH2 mRNA和蛋白的相对表达水平较阴性对照组明显下降(P<0.05),细胞对顺铂的敏感性显著增强,其半数抑制浓度(IC50值)与阴性对照组比较差异有统计学意义(P<0.05)。顺铂作用于转染let-7e mimics的A2780/CP细胞后,细胞的存活率较阴性对照组明显降低(P<0.05)。结论:let-7e在卵巢癌耐药细胞A2780/CP中异常低表达,上调其表达可部分逆转细胞耐药性。let-7e可能通过作用于靶蛋白EZH2参与卵巢癌细胞顺铂耐药的发生和发展。Objective：To investigate the expression of microRNA let-7e in cisplatin-re-sistant ovarian cancer cells and its mechanism. Methods：Stem-loop real-time PCR was used to detect the expression of let-7e in cisplatin-resistant ovarian cancer cell line A2780 / CP and its parent cell line A2780. The mRNA and protein levels of EZH2 in A2780 and A2780 / CP cells were detected by real-time PCR and Western blot assay,respectively. After transfection of let-7e mimics and negative control,the mRNA and protein levels of EZH2 in A2780 / CP cells were de-tected by real-time PCR and Western blot assay. MTT assay was used to analyze the half inhibi-tion concentration. The viability of A2780 / CP cells treated with cisplatin after transfection with let-7e mimics was determined by Trypan blue exclusion assay. Results：The expression of let-7e was significantly down-regulated in A2780 / CP cells compared with the parent cell line （ P 〈0. 05）. The mRNA and protein levels of EZH2 were up-regulated in A2780 / CP cells compared with A2780 cells （P〈0. 05）. After transfection of let-7e mimics,the mRNA and protein levels of EZH2 in A2780 / CP cells were significantly reduced. The IC50 of A2780 / CP cells transfect-ed with let-7e mimics was lower than that in the negative control（P〈0. 05）. Meanwhile,the vi- 〈br〉 anbeiglaittyiveofcAo2n7tr8o0l/CP cells transfected with let-7e mimics was significantly lower than that in the（P〈0. 05）. Conclusion：The expression of let-7e was down-regulated inA2780/CP cells. Up-regulation the expression of let-7e could partially reverse the cisplatin-re-ability of A2780 / CP cells transfected with let-7e mimics was significantly lower than that in the negative control （ P 〈 0. 05）. Conclusion： The expression of let-7e was down-regulated in A2780 / CP cells. Up-regulation the expression of let-7e could partially reverse the cisplatin-re-sistance. let-7e may play a vital role in drug resistance by targeting EZH2.

关 键 词：卵巢癌 耐药 顺铂 EZH2 EZH2 

分 类 号：R737.33[医药卫生—肿瘤]