骨化三醇、雷帕霉素及其联合使用对人子宫内膜癌Ishikawa细胞株的作用及机制研究

Effect of calcitriol,rapamycin and their combination on human endometrial carcinoma Ishikawa cells in vitro and its mechanism

目的观察骨化三醇、雷帕霉素及其联合使用对人子宫内膜癌Ishikawa细胞抑癌作用及其可能的作用机制。方法采用MTT法检测不同浓度的骨化三醇、雷帕霉素对人子宫内膜癌细胞株Ishikawa抑制率的影响;用倒置显微镜观察细胞形态学改变;流式细胞仪检测细胞增殖周期的变化;RT-PCR检测Akt、mTOR mRNA的表达;Western blot检测Akt、mTOR蛋白的表达。结果 10^(-7) mol·L^(-1)骨化三醇组和50.0 nmol·L^(-1)的雷帕霉素与人子宫内膜癌Ishikawa细胞株共培养48 h为二者抑癌效应适宜浓度和时间点。联合用药组对细胞的增殖抑制率均高于单独应用骨化三醇、雷帕霉素组,而Akt/mTOR信号物质表达量均明显降低(P<0.05)。结论骨化三醇联合雷帕霉素可协同抑制人子宫内膜癌细胞株Ishikawa的增殖,其机制可能是通过抑制PI3K/Akt/mTOR信号通路,从而发挥抑制子宫内膜癌增殖的作用。

To explore the effect of calcitriol,rapamycin and their combination administeredd to the human endometrial carcinoma Ishikawa cells and the underlying mechanism.Methods The growth inhibition ratio of the human endometrial carcinoma Ishikawa cells administered by calcitriol,rapamycin and their combination in vitro was observed by MTT.The morphological changes of the cells were observed using phase-contrast microscopy.The proliferation cycle was detected with flow cytometry.The expressions of Akt and mTOR mRNA and protein were assessed by RT-PCR and Western blot,respectively.Results It was a suitable concentration and time point of 10-7 mol·L-1 calcitriol,50.0 nmol·L-1 rapamycin and human endometrial carcinoma Ishikawa cells co-culture about 48 h.The inhibitory rate of the combination group was higher than that of the calcitriol and rapamycin group(P<0.05).However,the expressions of Akt and mTOR mRNA and proteins of the combination group were significantly lower than those of the calcitriol and rapamycin group alone(P<0.05).Conclusions The calcitriol,rapamycin and their combination with appropriate concentration and time may inhibit Ishikawa cell growth significantly,but the effect of their combination is better than that of the calcitriol and rapamycin alone.The mechanism may be related to the inhibition of the signaling pathway of PI3K/Akt/mTOR.