靶向PDCD5-Caspase-3融合基因过表达对HepG2肝癌细胞增殖与凋亡的影响  

Effect of PDCD5-Caspase-3 fusion gene overexpression on proliferation and apoptosis of HepG2 hepatoma cells

作　　者：胥丹 姜淮芜 蒲羽 Dan Xu;Huai-wu Jiang;Yu Pu(Southwest Medical University,Luzhou,Sichuan 646000,China;Department of Abdominal Surgery,Sichuan Mianyang 404 Hospital,Mianyang,Sichuan 621000,China)

机构地区：[1]西南医科大学,四川泸州646000 [2]四川绵阳404医院腹部外科,四川绵阳621000

出　　处：《中国现代医学杂志》2018年第24期22-26,共5页China Journal of Modern Medicine

摘　　要：目的探讨靶向PDCD5-Caspase-3融合基因过表达对Hep G2肝癌细胞体外增殖与凋亡的影响。方法利用基因重组技术构建PDCD5-Caspase-3融合基因,克隆到带有EGFP基因的GV358慢病毒表达载体,与辅助质粒共同转染293T细胞,通过同源重组,获得融合基因重组慢病毒PDCD5-Caspase-3。体外转染人肝癌Hep G-2细胞72 h后,Western bolt检测肝癌Hep G-2细胞中PDCD5蛋白和Caspase-3蛋白表达,CCK-8法检测PDCD5-Caspase-3融合基因过表达对Hep G-2肝癌细胞增殖的影响,流式细胞术观察其对细胞凋亡的影响。结果 PDCD5-Caspase-3融合基因过表达可增加Hep G-2肝癌细胞中PDCD5蛋白和Caspase-3蛋白合成(P<0.05);与对照组或空白组相比,转染72 h后实验组细胞增殖能力下降(P<0.05);与对照组或空白组比较,转染72 h后实验组细胞凋亡率增加(P<0.05)。结论促凋亡基因PDCD5和Caspase-3联合表达可抑制Hep G2肝癌细胞增殖,并促进其凋亡,可作为肝癌治疗潜在靶点。Objective To investigate the effect of PDCD5-Caspase-3 fusion gene overexpression on proliferation and apoptosis of human liver cancer HepG2 cell line. Methods PDCD5-Caspase-3 fusion gene was constructed and sub-cloned into GV358 eukaryotic expression vector. 293T cells were cotransfected with helper plasmid and virus. PDCD5-Caspase-3 fusion gene recombinant lentivirus was transfected into HepG2 hepatoma cells for 72 hours. Expression of PDCD5 and Caspase-3 protein was measured by Western blot. Cell proliferation and apoptosis were identified by CCK-8 and flowcytometry, respectively. Results PDCD5 and Caspase-3 were significantly increased in HepG2 hepatoma compared with control group （P 〈 0.05）. Cellular proliferation was decreased while apoptosis rate was increased 72 hours after transfection in transfection group （P 〈 0.05）. Conclusion Overexpression of PDCD5- Caspase-3 fusion gene inhibits cancer cell growth and may be a potential therapeutic target.

关 键 词：PDCD5 CASPASE-3 融合基因 细胞增殖 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]