miR-141靶向YAP1抑制肝癌细胞HCC-LM3增殖、迁移和侵袭

目的:探究microRNA-141(miR-141)对肝癌细胞HCC-LM3增殖、迁移、侵袭的影响。方法:通过荧光定量PCR(qPCR)检测肝癌细胞系(HepG2、Huh7、HCC-LM3)与肝上皮细胞THLE-3中miR-141的表达量;免疫印迹试验(Western blot)分析Yes相关蛋白1(YAP1)的表达量。采用噻唑蓝(MTT)实验分析过表达miR-141或沉默YAP1对HCC-LM3细胞增殖的影响,Transwell实验检测细胞的侵袭、迁移能力。采用生物信息学预测和双荧光素酶报告基因实验验证miR-141的靶基因。功能性实验检测过表达YAP1对miR-141调控的HCC-LM3细胞增殖、迁移、侵袭作用的影响。结果:qPCR和Western blot的结果表明,肝癌细胞系(HepG2、Huh7、HCC-LM3)中miR-141的表达量下调,YAP1的表达量上调;过表达miR-141和沉默YAP1可以抑制HCC-LM3细胞增殖、侵袭、迁移;生物信息学预测YAP1可能是miR-141的靶基因,双荧光素酶报告基因证实YAP1是miR-141的靶基因;过表达YAP1逆转miR-141对HCC-LM3细胞增殖、迁移、侵袭的抑制作用。结论:miR-141直接靶向YAP1抑制肝癌HCC-LM3细胞增殖、迁移、侵袭。

MiR-141 inhibits proliferation,migration and invasion of hepatocellular carcinoma cell line HCC-LM3 via directly targeting YAP1

Objective:To investigate the effect of microRNA-141 (miR-141) on the proliferation,invasion and migration of hepatocellular carcinoma cell line HCC-LM3.Methods:The expression of miR-141 in hepatocellular carcinoma cell lines (HepG2,Huh7,HCC-LM3) and hepatic epithelial cells THLE-3 were detected by real-time PCR (qPCR).The expression level of Yes-related protein 1(YAP1) was analyzed by Western blot.The effects of overexpression of miR-141 or silencing YAP1 on the proliferation of HCC-LM3 cells were analyzed by thiazolyl blue (MTT) assay.Transwell assay was used to detect the invasion and migration ability of cells.The targeted genes of miR-141 was predicted and verified by bioinformatics prediction and dual luciferase reporter gene experiments.Functional experiments examined the effect of overexpression of YAP1 on proliferation,migration,and invasion of HCC-LM3 cells regulated by miR-141.Results:The expression of miR-141 was down-regulated and the expression of YAP1 was up-regulated in hepatocellular carcinoma cell lines (HepG2,Huh7,HCC-LM3).Overexpression of miR-141 and silencing YAP1 inhibited HCC- LM3 cell proliferation,invasion,migration.Bioinformatics algorithms showed that YAP1 was likely to be a target gene of miR-141,and dual luciferase reporter gene confirmed YAP1 was a target gene of miR-141.Overexpression of YAP1 reversed the inhibitory effect of miR-141 on proliferation,migration and invasion of HCC-LM3 cells.Conclusion:miR-141 directly targets YAP1 to inhibit proliferation,migration and invasion of hepatocellular carcinoma HCC-LM3 cells.