抗肿瘤药物雷帕霉素调控子宫内膜癌细胞增殖、分化、凋亡的研究机制

摘要

Objective To investigate the effects of rapamycin on the proliferation, differentiation and apoptosis of endometrial carcinoma cells.Methods Cell proliferation was detected after 5 nmol/L, 10 nmol/L, 20 nmol/L, 40 nmol/L, 80 nmol/L, 160 nmol/L of rapamycin treated endometrial carcinoma HEC-1B cells for 24 h, 48 h, 72 h by CCK8, and IC50 was calculated;ALP mRNA expression was detected after 36nmol/L rapamycin treated HEC-1B cells for 0 d, 1 d, 3 d, 5 d, 7 d by RT-PCR;cell apoptosis and Cleaved Caspase3, mTOR, p-p70S6K protein expression was detected after 36nmol/L rapamycin treated HEC-1B for 48 h by flow cytometry and Western blot.Results With the increase of concentration and time, cell survival rate in 10 nmol/L, 20 nmol/L, 40 nmol/L, 80 nmol/L, 160 nmol/L rapamycin group were significantly lower than control group (P<0.01), according to the IC50 value than 36 nmol/L rapamycin was selected as follow-up research object;ALP mRNA expression after rapamycin treated HEC-1B cells for 1 d, 3 d, 5 d, 7 d were significantly higher than control group (P<0.01).Compared with the control group, the apoptosis rate and the expression of Cleaved Caspase3 protein were significantly increased, the expression of mTOR and p-p70S6K protein was significantly lower (P<0.01).Conclusion Rapamycin can significantly inhibit the proliferation of HEC-1B cells, induce cell differentiation and promote apoptosis, which may be related to the inhibition of mTOR/p70S6K signaling pathway.%目的 雷帕霉素对子宫内膜癌细胞增殖、分化、凋亡的影响及机制.方法 5 nmol/L、10 nmol/L、20 nmol/L、40 nmol/L、80 nmol/L、160 nmol/L的雷帕霉素作用于子宫内膜癌HEC-1B细胞24 h、48 h、72 h后,CCK8检测细胞增殖,计算IC50;36 nmol/L的雷帕霉素作用于HEC-1B细胞0 d、1 d、3 d、5 d、7 d后,RT-PCR检测细胞中ALP的mRNA表达;36 nmol/L的雷帕霉素作用于HEC-1B细胞48 h,流式细胞仪检测细胞凋亡,Western blot检测Cleaved caspase3、mTOR、p-p70S6K蛋白表达.结果 随着时间及浓度的增加,10 nmol/L、20 nmol/L、40 nmol/L、80 nmol/L、160 nmol/L的雷帕霉素组细胞存活率均显著低于对照组(P<0.01),根据IC50值选择36 nmol/L的雷帕霉素作为后续研究对象;ALP在1 d、3 d、5 d、7 d的mRNA表达均显著高于对照组(P<0.01);与对照组比较,细胞凋亡率及Cleaved caspase3蛋白表达均显著升高,mTOR、p-p70S6K蛋白表达显著下调(P<0.01).结论 雷帕霉素能显著抑制子宫内膜癌HEC-1B细胞增殖,诱导细胞分化及促进细胞凋亡,其机制与抑制mTOR/p70S6K信号通路有关.