恩度调控TGF-β/TβR/Smad2/3通路对大鼠HSC-T6细胞人肝癌HepG2细胞凋亡及迁移能力的影响

Influence of Endostar regulating TGF-β/TβR/Smad2/3 pathway on apoptosis and migration of rat HSC-T6 cells and hepatocellular carcinoma HepG2 cells

目的探讨恩度调控转化生长因子-β(transforming growth factor-β,TGF-β)/转化生长因子-β受体(TGF-βreceptor,TβR)/Smad2/3通路对大鼠HSC-T6细胞、肝癌HepG2细胞凋亡及迁移能力的影响。方法体外培养大鼠HSC-T6细胞、人肝癌HepG2细胞,均分为空白对照组、TGF-β1组(加入TGF-β1处理1 h)、TGF-β1+低剂量恩度组(加入TGF-β1处理1 h后加入20 pmol/L恩度处理24 h)、TGF-β1+中剂量恩度组(加入TGF-β1处理1 h后加入40 pmol/L恩度处理24 h)、TGF-β1+高剂量恩度组(加入TGF-β1处理1 h后加入80 pmol/L恩度处理24 h)。每组均设置3个复孔,且完成3次独立重复实验。采用Annexin V-FITC/PI双染法检测细胞凋亡率,采用划痕法检测细胞迁移率,采用免疫印迹法检测TβRⅠ、TβRⅡ、Smad2、Smad3蛋白表达量。结果与空白对照组相比,TGF-β1组的细胞凋亡率降低,细胞迁移率增加,TβRⅠ、TβRⅡ、Smad2/3蛋白表达量增加(P<0.05)。与TGF-β1组相比,TGF-β1+低、中、高剂量恩度组的细胞凋亡率增加,细胞迁移率降低,TβRⅠ、TβRⅡ、Smad2、Smad3蛋白表达量降低(P<0.05)。结论恩度可能通过下调TGF-β/TβR/Smad2/3通路,促进大鼠HSC-T6细胞、肝癌HepG2细胞凋亡,并抑制细胞迁移能力,从而发挥抗肝纤维化、肝癌作用。

Objective To investigate the influence of Endostar regulating transforming growth factor-β(TGF-β)/TGF-β receptor(TβR)/Smad 2/3 pathway on apoptosis and migration of rat hepatic stellate cells(HSC-T6 cells)and hepatocellular carcinoma HepG2 cells.Methods Rat HSC-T6 cells and hepatocellular carcinoma HepG2 cells were cultured in vitro,and divided into blank control group,TGF-β1 group,TGF-β1+low-dose Endostar group,TGF-β1+medium-dose Endostar group,and TGF-β1+high-dose Endostar group.The cell apoptosis rate,the cell migration rate and the protein expression levels of TβRⅠ,TβRⅡ,Smad2 and Smad3 were detected.Results Compared with the blank control group,the apoptosis rate was significantly decreased,the migration rate and the protein expression levels of TβRⅠ,TβRⅡ,Smad2 and Smad3 were significantly increased in TGF-β1 group(P<0.05).Compared with the TGF-β1 group,the apoptosis rate was significantly increased,the migration rate and the protein expression levels of TβRⅠ,TβRⅡ,Smad2 and Smad3 were significantly decreased in the TGF-β1+low-dose Endostar group,the TGF-β1+medium-dose Endostar group and the TGF-β1+high-dose Endostar group(P<0.05).Conclusion Endostar may promote the apoptosis of rat HSC-T6 cells and hepatocellular carcinoma HepG2 cells,and inhibit cell migration by down-regulating the TGF-β/TβR/Smad2/3 pathway,thereby exerting anti-hepatic fibrosis-hepatocellular carcinoma effect.