IGF-II促进人肝癌细胞MHCC97-H生长增殖作用的研究

摘要

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MH-CC97-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MH-CC97-H细胞形态发生一定改变；IGF-II时间依赖性地增加 MHCC 97- 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 effect of IGF -II on growth and proliferation in MHCC97 -H cells. Methods:Morphology of cells was observed under microscope. Relative viability( MTT-assay)of MHCC97-H cells was determined after application of IGF-II(25,50,100 ng/ml)at different time points(24,48,and 72h). Relative proliferation(BrdU-assay)of MHCC97-H cells was measured 48h after application of IGF-II. The clone formation efficiency of cells was determined by plate clone formation experiment. Expression of characteristic markers of prolifer-ation( PCNA )was identified with immunofluorescence. Furthermore,protein expression of PCNA was detected by Western blot. Results:MHCC97 -H cells induced by IGF -II showed a morphological alteration. MTT,BrdU and clone-formation assay suggested that growth and proliferation of MHCC97-H cells were increased by IGF-II,but no significant difference was found between IGF-II groups compared with control group(P>0. 05). Moreover,com-pared with the control,immunofluorescent expression of PCNA was evidently enhanced by IGF-II(P<0. 01). And protein expression of PCNA was significantly increased by IGF-II(P<0. 05). Conclusion:IGF-II might induce growth and proliferation and up-regulate PCNA expression trend in MHCC97-H cells.