RAD001对胃癌细胞株MKN 28、N 87生长的实验研究

目的探讨RAD001作用于胃癌细胞株MKN 28、N 87后细胞生长的影响及其作用机制。方法体外培养胃癌MKN 28、N 87细胞，给予RAD001干预后通过细胞计数、流式细胞术、Western blot电泳检测肿瘤细胞生长、细胞周期分布及mTOR、p70S6K、S6蛋白表达及磷酸化情况。结果细胞计数结果显示RAD001作用24?h即开始出现对细胞生长的抑制作用，作用具有时间依赖性。流式细胞术结果显示RAD001作用后两细胞株细胞周期均发生改变，停滞在G0 G1期细胞比例增加，作用亦具有时间依赖性，其中N 87相对敏感。Western blot结果显示RAD001作用24?h后两细胞株mTOR、p70S6K及 phosphate mTOR蛋白表达无改变，MKN 28 S6表达下降，phosphate p70S6K表达下调，phosphate S6表达缺失；N 87 S6无明显变化，phosphate p70S6K表达下调，phosphate S6表达明显下降。结论RAD001通过与mTOR的FRB相结合，从而抑制mTOR所调节的下游p70S6K S6蛋白通路，阻断了p70S6K及S6蛋白的磷酸化和蛋白质翻译进而调节着细胞周期以发挥抑制胃癌细胞生长的作用。

Effect of RAD001 on cell growth of gastric carcinoma cell lines MKN-28,N-87:an empirical study

Objective To investigate the effect of RAD001 on growth of gastric carcinoma cell lines MKN-28,N-87 and to investigate its possible mechanism.Methods Gastric carcinoma cell lines were treated by RAD001(20nM),then cell coulter, flow cytometry,and western-blot array were used to detect the growth,cell cycle and protein expression of tumor cells.Results After being treated by RAD001,gastric carcinoma cells were inhibited in a time-dependent manner.Percentage of Go phase was increased in the two cell lines.Also...