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目的:探讨IGF-Ⅱ在人肝癌细胞MHCC97-H生长增殖中的作用。方法:培养人肝癌细胞株MHCC97-H,IGF-Ⅱ(25,50,100ng/ml)作用于不同时间点(24、48、72h),观察各组细胞形态变化,噻唑兰(MTT)法分析细胞生长情况,Brdu法检测细胞增殖潜能,平板克隆实验观察并计算细胞克隆形成率,细胞免疫荧光及Western blot法观察细胞增殖核抗原(PCNA)表达,进一步分析细胞增殖情况。结果:IGF-Ⅱ可使MHCC97-H细胞形态发生一定改变;IGF-Ⅱ时间依赖性地增加MHCC9 7-H细胞存活率,但与对照组比较无显著差异(P>0.05);IGF-Ⅱ可促进MHCC97-H细胞增殖,但与对照组比较无显著差异(P>0.05);IGF-Ⅱ(50ng/ml)对MHCC97-H细胞的克隆形成有一定的促进作用,但与对照组比较无显著差异(P>0.05);与对照组相比,IGF-Ⅱ(50ng/ml)组,MHCC97-H细胞中PCNA荧光表达和蛋白表达显著升高(P<0.05)。结论:IGF-Ⅱ有诱导人肝癌细胞MHCC97-H生长增殖的趋势,亦可促进PCNA表达上调。
Objective: To investigate the role of IGF-Ⅱ in the growth and proliferation of human hepatoma cell line MHCC97-H. Methods: The human hepatocellular carcinoma cell line MHCC97-H and IGF-Ⅱ (25, 50 and 100 ng / ml) were cultured at different time points (24, 48 and 72 hours) Brdu method was used to detect the cell proliferation potential. The clone formation assay was performed by plate clone assay. The expression of PCNA was observed by immunofluorescence and Western blot. The cell proliferation was further analyzed. Results: The morphology of MHCC97-H cells was changed by IGF-Ⅱ. IGF-Ⅱ could increase the survival rate of MHCC9 7-H cells in a time-dependent manner, but there was no significant difference compared with the control group (P> 0.05) (P> 0.05). IGF-Ⅱ (50ng / ml) could promote the cloning of MHCC97-H cells, but there was no significant difference compared with the control group (P> 0.05). Compared with the control group, the expression of PCNA and protein in MHCC97-H cells in IGF-Ⅱ (50ng / ml) group were significantly increased (P <0.05). Conclusion: IGF-Ⅱ can induce the proliferation and proliferation of human hepatoma cell line MHCC97-H, and also promote the up-regulation of PCNA expression.