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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。
DEP domain containing protein 7 (DEP domain containing 7) is obtained from data mining of gene chip, and gene expression profile suggests that it is selectively expressed only in liver tissue. However, it is related to the involvement of DEPDC7 in human hepatocellular carcinoma cells and their molecular mechanisms Less research reports. In this study, DEPDC7 lentiviral vector was constructed and infected with human hepatocellular carcinoma cell line HepG2 by using RNAi technique. The interference effect of DEPDC7 was detected by RT-q PCR and Western blot. Cell proliferation was detected by MTT colorimetric assay and clonogenic assay. Flow cytometry was used to detect cell cycle changes. Transwell chamber was used to detect cell invasion and migration. The results showed that the constructed DEPDC7 lentiviral vector could effectively interfere with the expression of DEPDC7 mRNA and protein (P <0.05). In addition, silencing of DEPDC7 could effectively promote cell cycle transition from G1 phase to S phase, and significantly enhanced cell proliferation and invasion and migration (P <0.05). This study suggests that low expression of DEPDC7 in HepG2 cells can effectively increase cell proliferation, colony formation and invasion and migration, indicating the direction of DEPDC7 transcriptional regulation.