miR-654-3p通过靶向抑制TOP2A增加宫颈癌细胞对顺铂的敏感性  

miR-654-3p increases the sensitivity of cervical cancer cells to cisplatin by targeting

作　　者：魏晶[1] 高玉华 李卓[1] TOPA WEI Jing;GAO Yu-hua;LI Zhuo(Gynecological Radiotherapy,Liaoning Cancer Hospital,Shenyang 110042,China)

机构地区：[1]辽宁省肿瘤医院妇科放疗,辽宁沈阳110042

出　　处：《解剖科学进展》2024年第5期548-551,共4页Progress of Anatomical Sciences

基　　金：新锐肿瘤支持治疗课题研究项目(cphcf-2022-174)。

摘　　要：目的探究miR-654-3p对顺铂耐药的宫颈癌Hela/DDP细胞对顺铂敏感性的影响,并进一步分析其分子机制。方法生物信息学预测TOP2A mRNA 3′-UTR的miR-654-3p潜在结合位点,双荧光素酶报告基因实验验证。RT-qPCR和Western blot方法检测Hela和Hela/DDP细胞中miR-654-3p和TOP2A表达情况以及过表达miR-654-3p对Hela/DDP细胞中TOP2A表达的影响。将TOP2A过表达质粒转染过表达miR-654-3p的Hela/DDP细胞,经顺铂处理后,CCK-8和流式细胞术分别检测Hela/DDP细胞的细胞活力和凋亡率。结果TOP2A mRNA 3′-UTR存在miR-654-3p的潜在结合位点,且miR-654-3p与TOP2A mRNA 3′-UTR结合。Hela/DDP细胞中miR-654-3p表达水平较Hela细胞下调,TOP2A表达水平较Hela细胞上调。过表达miR-654-3p降低Hela/DDP细胞中TOP2A表达。过表达TOP2A逆转了过表达miR-654-3p对Hela/DDP细胞中TOP2A表达的下调作用。过表达miR-654-3p降低顺铂处理的Hela/DDP细胞的细胞活力,增加Hela/DDP细胞的凋亡率;过表达TOP2A逆转了过表达miR-654-3p对顺铂处理的Hela/DDP细胞的细胞活力的下调作用和凋亡率的上调作用。结论MiR-654-3p通过靶向抑制TOP2A表达增加顺铂耐药的宫颈癌细胞对顺铂的敏感性。Objective To explore the effect of miR-654-3p on the sensitivity of cisplatin-resistant cervical cancer Hela/DDP cells to cisplatin,and further analyze its molecular mechanism.Methods Bioinformatics predicts the potential binding site of miR-654-3p of TOP2A mRNA 3′-UTR and verifies the binding of them by double luciferase reporter gene experiment.RT-qPCR and Western blot were used to detect the expressions of miR-654-3p and TOP2A in Hela and Hela/DDP cells,and the effect of overexpression of miR-654-3p on TOP2A expression in Hela/DDP cells.TOP2A overexpression plasmid was transfected into Hela/DDP cells expressing miR-654-3p.After cisplatin treatment,the cell viability and apoptosis rate of Hela/DDP cells were detected by CCK-8 and flow cytometry respectively.Results There was a potential binding site of miR-654-3p in TOP2A mRNA 3′-UTR,and miR-654-3p binded to TOP2A mRNA 3′UTR.The expression level of miR-654-3p in Hela/DDP cells was down-regulated and the expression level of TOP2A was up-regulated.Overexpression of miR-654-3p decreased the expression of TOP2A in Hela/DDP cells.Overexpression of TOP2A reversed the down-regulation effect of overexpression of miR-654-3p on TOP2A expression in Hela/DDP cells.Overexpression of miR-654-3p decreased the cell viability of Hela/DDP cells treated with cisplatin and increased the apoptosis rate of Hela/DDP cells.Overexpression of TOP2A reversed the down-regulation of miR-654-3p on cell viability and up-regulation of apoptosis rate of Hela/DDP cells treated with cisplatin.Conclusion MiR-654-3p could increase the sensitivity of cisplatin-resistant cervical cancer cells to cisplatin by targeted inhibition of TOP2A expression.

关 键 词：miR-654-3p 宫颈癌 顺铂 敏感性 TOP2A 

分 类 号：R737.33[医药卫生—肿瘤]