SOX7基因对肝癌细胞凋亡、氧化应激及Wnt/β-catenin信号通路的影响

Effect of SOX7 gene on the apoptosis,oxidative stress and Wnt/β-catenin signaling pathway in liver cancer cells

目的:探讨过表达SOX7对肝癌细胞凋亡、活性氧(ROS)水平及Wnt/β-catenin信号通路的影响。方法:人肝癌Huh-7细胞分为空白组、空质粒pcDNA3.1组(Vector组)和pcDNA3.1-SOX7重组质粒组(pcDNA3.1-SOX7组)。转染48h,AnnexinV-FITC/PI双染法细胞凋亡试剂盒、DCFH-DA法及Westernblotting分别检测细胞凋亡率、ROS含量及SOX7、β-catenin、cyclinD1和cleaved-caspase3蛋白表达。MTT法检测pcDNA3.1转染24h、48h和72h的细胞活力。结果:pcDNA3.1-SOX7重组质粒组SOX7蛋白表达明显高于空白组(P<0.05)。与空白组比较,pcDNA3.1-SOX7组细胞活力及β-catenin和cyclinD1蛋白表达均明显降低,细胞凋亡率、ROS含量及cleaved-caspase3蛋白表达明显升高(P<0.05)。结论:过表达SOX7可诱导肝癌细胞凋亡,凋亡机制与细胞内ROS水平提高及Wnt/β-catenin信号通路抑制有关。

Objective: To investigate the effects of overexpression of SOX7 gene on apoptosis,ractive oxygen species(ROS) level and Wnt/β-catenin signaling pathway in hepatocellular carcinoma(HCC) cells. Methods: The human hepatoma Huh-7 cells were divided into the blank group,blank plasmid pcDNA3.1 group(vector group) and pcDNA3.1-SOX7 recombinant plasmid group(pcDNA3.1-SOX7 group).The apoptosis rate,ROS content,and expression levels of SOX7,β-catenin,cyclin D1 and cleaved-caspase 3 protein were detected by Annexin V-FITC/PI double staining,DCFH-DA assay and Western blotting at 48 h after transfection,respectively.The cell viability was detected using MTT assay after 24 h,48 h and 72 h of pcDNA3.1 transfecting. Results: The expression level of SOX7 protein in pcDNA3.1-SOX7 recombinant plasmid group was significantly higher than that in blank group( P <0.05).Compared with the blank group,the cell viability and expression levels of β-catenin and cyclin D1 protein in pcDNA3.1-SOX7 group significantly decreased,while the apoptosis rate,ROS content and cleaved-caspase 3 protein expression level significantly increased( P <0.05). Conclusions: The overexpression of SOX7 can induce the apoptosis of HCC cells.The mechanism of apoptosis is related to the increasing of ROS level and inhibition of Wnt/β-catenin signaling pathway.