shRNA介导DEPDC7基因沉默对肝癌HepG2细胞增殖、侵袭及迁移能力影响的研究

Effects of sh RNA-mediated Knockdown of DEPDC7 on Proliferation, Migration and Invasion of Human Hepatoma Cell HepG2

功能未知基因DEPDC7(DEP domain containing 7)是从基因芯片数据挖掘得到的,基因表达谱提示只在肝组织中选择性高表达,但是关于DEPDC7在肝癌细胞内参与的生命活动及其分子机制的相关研究报道较少。该研究应用RNAi技术构建DEPDC7慢病毒载体并感染人肝癌细胞株HepG2,利用RT-q PCR和Western blot方法检测DEPDC7干扰效果。运用噻唑蓝(MTT)比色法和克隆形成方法检测细胞增殖能力,流式细胞术检测细胞周期变化,Transwell小室检测细胞侵袭迁移能力。结果显示,成功构建的DEPDC7慢病毒载体可以有效干扰DEPDC7 m RNA和蛋白的表达(P<0.05)。此外,DEPDC7被沉默后,可以有效促进细胞周期从G1期向S期转变,细胞增殖和侵袭迁移能力均显著提高(P<0.05)。该研究提示,肝癌细胞HepG2中DEPDC7低表达能有效提高细胞增殖、克隆形成和侵袭迁移能力,为后续研究DEPDC7转录调控机制等指明了方向。

Function-unknown gene DEPDC7（DEP domain containing 7） was obtained by gene chip data mining with highly selective expression in liver tissue. However, its role and molecular mechanism in hepatic cellular carcinoma are still unclear. In this study, we utilized liver cancer cell line HepG2 to construct DEPDC7 knockdown cell strain by RNAi. Moreover, we analyzed the m RNA and protein expression levels of DEPDC7 by RT-q PCR and Western blot. Cell proliferation and cell cycle were analyzed by methyl thiazol tetrazolium（MTT）, colony formation experiment and flow cytometry（FACS）. Cell motility and invasiveness were assayed by Matrigel migration and invasion assay. The results showed that DEPDC7-sh RNA effectively inhibited the expression of DEPDC7 at both m RNA and protein levels（ P〈0.05）. Knockdown of DEPDC7 led to increased proliferation, migration and invasion（ P〈0.05）. Our results suggested that sh RNA-mediated knockdown of DEPDC7 significantly promoted cell proliferation, colony formation, migration and invasion. Moreover, this research pointed out the direction of future research on the mechanism of transcription and regulation of DEPDC7.