金雀异黄酮与芹菜素的雌激素样作用体外研究

目的:用雌激素受体(estrogen receptor,ER)表达阳性细胞株人乳腺癌细胞MCF-7,检测金雀异黄酮(genistein)与芹菜素(apigenin)的雌激素样活性大小。方法:用MTT法研究金雀异黄酮与芹菜素对体外培养MCF-7细胞增殖的影响;Real-time RT-PCR法检测其对ERα,ERβ,PR,PS2 mRNA表达水平的影响。结果:金雀异黄酮与芹菜素促进MCF-7增殖;金雀异黄酮的1×10-10mol.L-1组显著增加ERαmRNA的表达水平,是对照组的17.76倍,是阳性对照E2组的1.75倍。芹菜素显著促进了PR mRNA表达水平,是对照组4.57倍,并达到E2组1.11倍。两者在不同浓度对ERα,ERβ,PR,PS2 mRNA中的一个或几个均有不同的促进作用。结论:金雀异黄酮与芹菜素均有明显的雌激素样作用,虽对于不同的雌激素应答基因(ERα,ERβ,PR,PS2 mRNA)表现出不同的促进表达作用,但金雀异黄酮更加强烈。提示植物雌激素发挥雌激素样作用的信号途径与雌激素信号途径并不完全相同;黄酮类药物的B环位置是雌激素样活性的关键位点之一,异黄酮(B环处于3位时)的雌激素样活性大于黄酮(B环处于2号位)。

Study on estrogenic effect of genistein and apigenin in vitro

Objective: To detect the estrogenic activity of genistein and apigenin with ER-positive cell line MCF-7 human breast cancer cells.Method: MTT method was adopted to study the impact of genistein and apigenin on MCF-7 proliferation in vitro.Real-time RT-PCR method was used to detect their impact on ERα,ERβ,PR and PS2 mRNA expression levels.Result: Genistein and apigenin promoted the proliferation of MCF-7.Genistein 1×10-10 mol·L-1 group showed a significant increase in the expression of ERα mRNA levels or a 17.76 times more than the control group and a 1.75 times more than the E2 group.Apigenin notably promoted the PR mRNA expression or a 4.57 times more than the control group and a 1.11 times more than the E2 group.Both of them had different effect in promoting ERα,ERβ,PR or PS2 mRNA.Conclusion: Both genistein and apigenin have a strong estrogen-like effect.Although they have different effect in promoting estrogenic response genes(such as ERα,ERβ,PR and PS2 mRNA),genistein shows a stronger activity than apigenin.It also suggests that the signaling pathways of phytoestrogens showing estrogen-like effect are not completely identical with estrogen pathways.The B-cycle position of flavonoids is one of the key sites to estrogen-like activity,and isoflavones(cycle B on site 3) show stronger estrogen-like activity than flavones(B-cycle lies in site 2).