槲皮素联合孕激素对人子宫内膜癌HEC-1-B细胞增殖、细胞周期及凋亡的影响

Study on effects of quercetin and progesterone on proliferation, cell cycle and apoptosis of human endometrial carcinoma HEC-1-B cells

目的探讨槲皮素联合孕激素对人子宫内膜癌HEC-1-B细胞体外增殖、细胞周期分布情况、凋亡的影响及其可能机制。方法槲皮素、孕酮单独及联合作用于HEC-1-B细胞后,运用MTT比色法观察各组对细胞增殖的作用;流式细胞仪检测各组细胞周期分布及细胞凋亡情况,Western Blot实验检测HEC-1-B细胞Cyclin D1、P21、Bcl-2与Bax的表达。结果槲皮素对HEC-1-B细胞的生长抑制率随药物浓度和作用时间的增加而增大,联合用药组的抑制作用更为明显(P<0.01)。流式细胞仪分析显示槲皮素将人子宫内膜癌HEC-1-B细胞阻滞于G_(0)/G_(1)期,槲皮素作用48 h后人子宫内膜癌HEC-1-B细胞凋亡率明显增加,且随药物浓度的增加而增大。联合用药组的作用更加显著。Western Blot结果显示,与对照组相比,槲皮素100、200μmol/L组及单独孕酮组作用48 h后,Cyclin D1、Bcl-2的蛋白表达降低,P21、Bax的蛋白表达升高。与孕酮组相比,槲皮素与孕酮联合作用可使Cyclin D1、Bcl-2的蛋白表达明显降低,P21、Bax的蛋白表达显著升高。结论槲皮素联合孕激素能明显抑制HEC-1-B细胞增殖,促进其凋亡,其机制可能与调控Cyclin D1、P21、Bcl-2与Bax的表达相关。

Objective To investigate the effect of quercetin combined with progesterone on the proliferation,cell cycle distribution and apoptosis of human endometrial cancer HEC-1-B cells in vitro and its possible mechanism.Methods The effect of quercetin and/or progesterone on cell proliferation,cell cycle and apoptosis of HEC-1-B cell was separately studied by MTT and flow cytometry.The expression of Cyclin D1,P21,Bcl-2 and Bax in HEC-1-B cells was detected by Western blot.Results The growth inhibition of quercetin on HEC-1-B was concentration-dependent and time-dependent and the united group presenteda more prominent(P<0.01).Flow cytometry analysis showed that quercetin blocked human endometrial cancer HEC-1-B cells in G_(0)/G_(1) period.After 48 h of quercetin treatment,the apoptosis rate of human endometrial cancer HEC-1-B cells increased significantly,and the united group presenteda more prominent.Western Blot results showed that,compared with the control group,the quercetin 100,200μmol/L group and the progesterone alone group after 48 hours of treatment,the protein expression of Cyclin D1 and Bcl-2 decreased,and the protein expression of P21 and Bax increased.Compared with the progesterone group,the combined effect of quercetin and progesterone can significantly reduce the protein expression of Cyclin D1 and Bcl-2,and significantly increase the protein expression of P21 and Bax.Conclusion Quercetin combine with progesterone can significant inhibit HEC-1-B cell proliferation and promote cells apoptosis,which may be related to the regulation of Cyclin D1,P21,Bcl-2 and Bax expression.