雷帕霉素对p70S6K-siRNA转染的 EC9706细胞增殖的影响

摘要

目的：观察雷帕霉素对p70S6K-siRNA转染的人食管鳞状细胞癌EC9706细胞增殖的影响。方法：用p70S6K-siRNA转染EC9706细胞，分别作用24、48 h后，采用RT-PCR和Western blot检测p70S6K mRNA及蛋白的表达情况；收集转染p70S6K-siRNA 24 h和未转染的EC9706细胞，加入0、25、50、100、150、250、500和1500 nmol/L的雷帕霉素处理48 h，采用MTT法检测细胞存活率。结果：p70S6K-siRNA转染后，EC9706细胞中p70S6K mRNA及蛋白的表达均明显降低（mRNA：F＝57．825；蛋白：Fp70S6K＝268．897，Fp-p70S6K ＝35．584，P均＜0．05）。随雷帕霉素浓度的增加，未转染和转染组EC9706细胞存活率均降低；与未转染组相比，转染组细胞存活率更低（ F剂量＝664．919， F组间＝1958．000，P均＜0．05。）结论：p70S6K-siRNA能增强雷帕霉素对EC9706细胞增殖的抑制作用。%Aim:To study the inhibition effects of rapamycin on cell proliferation after the expression of p 70S6K inter-fered by p70S6K-siRNA in esophageal squamous cell cacinoma EC 9706 cells .Methods:After p70S6K-siRNA being trans-ferred into EC9706 cells for 24 and 48 h,respectively, the expressions of p70S6K mRNA and protein were detected by RT-PCR and Western blot.The EC9706 cells transfected by P70S6K-siRNA for 24 h (the untransfected cells were control ), and then they were treated with 0,25,50,100,150,250,500 and 1 500 nmol/L rapamycin for 48 h,and then MTT assay was used to detect the cells'survival rate.Re sults:The mRNA and protein levels of p70S6K were decreased significantly in EC9706 cells treated with p70S6K-siRNA (mRNA:F=57.825,P<0.05;protein level: Fp70S6K =268.897,Fp-p70S6K =35.584,both P<0.05).The survival rate of cells was significantly decreased with the increase of the dose of rapamycin and after cells treated with p70S6K-siRNA (Fdose =664.919,Fgroup =1 958.000,both P<0.05).Conclusion: p70S6K-siRNA can improve the inhibition effects of rapamycin on proliferation of EC 9706 cells.