载脂蛋白B在肝细胞癌发生发展中的作用及机制研究  

The role and mechanism of apolipoprotein B in the development and progression of hepatocellular carcinoma

作　　者：刘媛 倪红妙 田华华 LIU Yuan;NI Hongmiao;TIAN Huahua(Hangzhou Ninth People's Hospital,Zhejiang 311225,China)

机构地区：[1]浙江省杭州市第九人民医院,杭州311225

出　　处：《浙江创伤外科》2023年第7期1202-1206,共5页Zhejiang Journal of Traumatic Surgery

基　　金：2019年浙江省杭州市卫生科技计划一般(B类)项目(OO20190898)。

摘　　要：目的观察载脂蛋白B(APOB)对肝细胞癌的增殖、迁移的影响及其相关作用机制研究。方法应用RNA干扰技术下调了人肝癌细胞株HepG2中的APOB表达,作为siRNA-APOB组,设计无关序列作为siRNA-NC组。依次应用qPCR验证干扰并筛选出最佳序列;CCK-8法及细胞划痕实验分别验证APOB对肝细胞癌增殖、迁移的影响;Western blot法检测干扰APOB后相关蛋白水平变化。结果qPCR验证siRNA-APOB抑制HepG2细胞中APOB表达成功。与siRNA-NC组比较,siRNA-APOB组细胞活力显著提高(P<0.01),即细胞增殖能力增强。与siRNA-NC组比较,siRNA-APOB组细胞愈合率显著提高(P<0.01),即迁移能力增强。与siRNA-NC组比较,siRNA-APOB组p38、LEF1、MITF、β-catenin蛋白表达水平均增加,其中LEF1、MITF、β-catenin蛋白表达水平增加具有统计学意义(P<0.05)。结论下调APOB表达可以促进HepG2细胞的增殖及迁移,还能够增加MITF、LEF1、β-catenin蛋白表达水平,其可能是通过促进Wnt/β-catenin/MITF/LEF1信号通路的激活实现。Objective To observe the effect of apolipoprotein B(APOB)on the proliferation and migration of hepatocellular carcinoma and its related mechanism of action.Methods RNA interference technology was applied to down-regulate APOB expression in human hepatocellular carcinoma cell line HepG2 as siRNA-APOB group and design irrelevant sequences as siRNA-NC group.The interference was verified by qPCR and the best sequence was screened;the effect of APOB on the proliferation and migration of hepatocellular carcinoma was verified by CCK-8 and cell scratch assay,respectively.The changes of related protein levels after interference with APOB were detected by Western blot.Results qPCR verified that siRNA-APOB inhibited APOB expression in HepG2 cells successfully.Compared with the siRNA-NC group,the cell viability of the siRNA-APOB group was significantly improved(P<0.01).The cell proliferation ability was increased.Compared with the siRNA-NC group,the cell healing rate was significantly improved(P<0.01)in the siRNA-APOB group.The migration ability was increased.Compared with the siRNA-NC group,the p38,LEF1,MITF andβ-catenin protein expression levels were improved in the siRNA-APOB group,among which the addition of LEF1,MITF andβ-catenin protein expression levels was statistically significant(P<0.05).Conclusion Downregulation of APOB expression promoted the proliferation and migration of HepG2 cells,and also increase the expression levels of MITF,LEF1,andβ-catenin proteins,which may be achieved by promoting the activation of Wnt/β-catenin/MITF/LEF1 signaling pathway.

关 键 词：载脂蛋白B(APOB) 肝细胞癌 RNA干扰技术 增殖 迁移 作用机制 

分 类 号：R735.7[医药卫生—肿瘤]