雌马酚对人卵巢癌SKOV-3细胞增殖的抑制作用及其机制研究

目的观察雌马酚(equol)对人卵巢癌SKOV-3细胞增殖的抑制作用,并探讨其作用的分子机制。方法用不同浓度(10-8、10-7、10-6、10-5、5×10-5、10-4mol/L)雌马酚处理SKOV-3细胞24、48、72h后,及同时用10μmol/LERK通路抑制剂U0126或雌激素受体抑制剂ICI182,780处理1h后再用50μmol/L雌马酚处理2h,测定各组SKOV-3细胞存活率,用Westernblotting检测总ERK和磷酸化ERK1/2(p-ERK)的蛋白表达水平。结果各实验组SKOV-3细胞存活率随着雌马酚作用浓度和时间的增加而降低,总ERK蛋白表达量不变,p-ERK的表达量随雌马酚处理浓度的增加而逐渐降低。U0126、ICI182,780单独使用或与雌马酚联合使用均能够降低p-ERK的蛋白表达。结论雌马酚显著抑制人卵巢癌SKOV-3细胞的增殖,其机制是通过下调磷酸化ERK蛋白的表达发挥作用。

THE INHIBITORY EFFECT OF EQUOL ON HUMAN OVARIAN CARCINOMA CELL SKOV-3 PROLIFERATION AND ITS MECHANISM

Objective To investigate the effect of equol on ERK mediated signal transduction pathway and to clarify its mechanism of proliferation inhibition on human ovarian carcinoma cell line SKOV-3. Method SKOV-3 cells were treated with 10~(-8), 10~(-7), 10~(-6), 10~(-5), 5×10~(-5), 10~(-4) mol/L equol for 24, 48 and 72h. After pretreatment with 10 nmoVL U0126 an ERK signaling pathway inhibitor or ICI182, 780, estrogen receptor inhibitors for lh, then treatment with 50 nmol/L equol for 2h, the cell viability was examined and the expressions of ERK and p-ERK protein were determined using Western blotting. Results Equol was demonstrated to inhibit SKOV-3, proliferation time- and dose-dependently. The expression of p-ERK protein was decreased in dose-dependent manner, while the expression of total ERK was unchanged. Both the single use of U0126, or ICI182, 780, and combined with equol could decrease the expression of p-ERK protein. Conclusion Equol could inhibit proliferation in ovarian carcinoma cell lines SKOV-3. Its inhibitory effect appears to be due to down-regulation of p-ERK protein.