瑞戈非尼通过miR-122调控人肝癌SMMC-7721细胞的增殖和凋亡  

Regorafenib regulates the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells through miR-122

作　　者：陈巍 韩峥 邹艳丽 黄莎莎 田霞 CHEN Wei;HAN Zheng;ZOU Yanli;HUANG Shasha;TIAN Xia(Department of Gastroenterology,Wuhan Third Hospital,Wuhan 430062,Hubei,China)

机构地区：[1]武汉市第三医院消化内科,湖北武汉430062

出　　处：《中国肿瘤生物治疗杂志》2021年第8期796-802,共7页Chinese Journal of Cancer Biotherapy

基　　金：湖北省中央引导地方科技发展专项资助(No.2019ZYYD067)。

摘　　要：目的:探究瑞戈非尼(regorafenib,Rego)对人肝癌SMMC-7721细胞增殖、凋亡的影响及其可能的机制。方法:将SMMC-7721细胞分为对照组及Rego组,分别用0、10μmol/L Rego处理48 h后,流式细胞术检测各组细胞凋亡率,qPCR检测细胞中miR-122的表达。采用脂质体转染的方法将合成的hsa-miR-122-5p模拟物转染SMMC-7721细胞构建miR-122过表达的overExp-miR-122细胞,并将细胞分为对照组、Rego组、overExp-NC组、overExp-NC+Rego组、overExp-miR-122组及overExp-miR-122+Rego组,采用MTT法检测细胞活性,流式细胞术检测细胞凋亡率、WB法检测细胞中Bcl2、cleaved caspase-3、RAS、RAF1、p-ERK1蛋白表达水平。结果:与对照组相比,Rego处理后细胞凋亡率显著升高(P<0.05),且miR-122表达量显著上升(P<0.01);与overExp-NC组比较,overExp-miR-122组细胞增殖抑制率、凋亡率和cleaved caspase-3表达均显著升高(均P<0.01),RAS蛋白表达显著下降(P<0.05),Bcl2、RAF、p-ERK1蛋白表达均显著下降(均P<0.01);与overExp-miR-122组相比,overExp-miR-122+Rego组细胞中各检测指标变化进一步显著增加(P<0.01)。结论:Rego可抑制SMMC-7721细胞增殖、促进凋亡,其作用可能与调控miR-122、凋亡相关因子的表达和抑制RAS/RAF/ERK信号通路有关。Objective:To explore the effects of regorafenib(Rego)on proliferation and apoptosis of human liver cancer SMMC-7721 cells and the possible mechanism.Methods:SMMC-7721 cells were divided into control group and Rego group(10μmol/L Rego).After the treatment for 48 h,Flow cytometry was used to detect cell apoptosis rate,and qPCR was used to determine the level of miR-122 in two groups of cells.hsa-miR-122-5p mimics were transfected into SMMC-7721 cells by lipofection to construct the miR-122 overexpressing cell line(overExp-miR-122).Then,the cells were divided into control group,Rego group,overExp-NC group,overExp-NC+Rego group,overExp-miR-122 group and overExp-miR-122+Rego group.MTT and Flow cytometry were used to detect cell viability and apoptosis rate,respectively.The protein expression levels of Bcl2,cleaved caspase-3,RAS,RAF1 and p-ERK1 in cells were detected by WB assay.Results:Compared with control group,the cell apoptosis rate was significantly increased after Rego treatment(P<0.05),and miR-122 expression level was significantly increased in Rego group(P<0.01).Compared with overExp-NC group,the proliferation inhibition rate and apoptosis rate in the cells of overExp-miR-122 group were significantly increased(P<0.01),the expression level of cleaved caspase-3 was significantly upregulated,while the protein expressions of Bcl2,RAS,RAF and p-ERK were significantly decreased(P<0.05 or P<0.01).Compared with overExp-miR-122 group,the changes in above detected indicators of overExp-miR-122+Rego group were more obvious(P<0.01).Conclusion:Regorafenib can inhibit the proliferation and promote apoptosis of SMMC-7721 cells,which may be achieved by regulating the expression of miR-122,thereby regulating the expression of apoptosis related factors and inhibiting RAS/RAF/ERK signaling pathway.

关 键 词：肝癌 SMMC-7721细胞 瑞戈非尼 MIR-122 凋亡 增殖 RAS/RAF/ERK信号通路 

分 类 号：R735.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]