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

摘要:

目的 探讨靶向PDCD5-Caspase-3 融合基因过表达对HepG2 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HepG-2 细胞72 h 后，Western bolt 检测肝癌HepG-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HepG-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HepG-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HepG2 肝癌细胞增殖，并促进其凋亡，可作为肝癌治疗潜在靶点。

Abstract:

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.