重组人生长激素对肝癌细胞MHCC-97H的作用及机制

Effects of human recombinant growth hormone on hepatocellular carcinoma cell line MHCC-97H and underlying mechanism

目的探讨重组人生长激素(rhGH)对肝癌细胞MHCC-97H的作用及机制。方法以无血清培养液诱导肝癌细胞MHCC-97H凋亡,再以浓度为0μg/L(GH0组)、5μg/L(GH5组)、25μg/L(GH25组)、50μg/L(GH50组)、100μg/L(GH100组)和500μg/L(GH500组)的rhGH完全培养液进行培养,MTT法观察MHCC-97H细胞生长情况,流式细胞技术检测细胞凋亡率、细胞周期和细胞增殖指数,RT-PCR和Western blotting法分别检测凋亡相关基因Bax和Bcl-2 mRNA和蛋白表达情况。结果与GH0组比较,GH25组、GH50组、GH100组和GH500组MHCC-97H细胞增殖明显,S期细胞百分比和细胞增殖指数显著增加(P<0.05);GH25组、GH50组和GH100组细胞凋亡率显著降低(P<0.05),Bax mRNA和蛋白表达显著降低(P<0.05),Bcl-2 mRNA和蛋白表达显著增加(P<0.05)。结论 rhGH体外显著促进肝癌细胞MHCC-97H增殖,并通过调节凋亡相关基因Bax和Bcl-2的表达来抑制无血清培养液诱导的细胞凋亡。

Objective To investigate the effects of recombinant human growth hormone（rhGH） on hepatocellular carcinoma cell line MHCC-97H,and explore the underlying mechanism.MethodsApoptosis of hepatocellular carcinoma cell line MHCC-97H was induced by serum free culture fluid,and MHCC-97H cells were then cultured with 0 μg/L rhGH（GH0 group）,5 μg/L rhGH（GH5 group）,25 μg/L rhGH（GH25 group）,50 μg/L rhGH（GH50 group）,100 μg/L rhGH（GH100 group） and 500 μg/L rhGH（GH500 group）,respectively.The growth of MHCC-97H cells was observed by MTT method,cell apoptosis rate,cell cycle and cell proliferation index were determined by flow cytometry,and RT-PCR and Western blotting were employed to detect the expression of Bax and Bcl-2 mRNA and protein,respectively.ResultsCompared with GH0 group,the proliferation of MHCC-97H cells,the percentages of cells in S phase and proliferation indexes significantly increased in GH25 group,GH50 group,GH100 group and GH500 group（P0.05）.Cell apoptosis rates in GH25 group,GH50 group and GH100 group significantly decreased（P0.05）,the expression of Bax mRNA and protein significantly decreased and that of Bcl-2 mRNA and protein significantly increased（P0.05）.ConclusionrhGH can stimulate the in vitro proliferation of hepatocellular carcinoma cell line MHCC-97H,the mechanism of which is closely related to the expression of Bax and Bcl-2 genes.