异补骨脂素的植物雌激素作用及其机制的探讨

目的利用雌激素受体(estrogenic receptor,ER)阳性乳腺癌T47D细胞和ER阴性乳腺癌MDA-MB231细胞观察异补骨脂素(isopsoralen)的雌激素样作用,并探讨其可能的作用靶点和机制。方法以ER拮抗剂ICI182,780为工具药,采用MTT细胞增殖实验观察异补骨脂素对T47D和MDA-MB231细胞增殖的影响;通过流式细胞术检测细胞周期分布的变化。同时,利用实时荧光定量PCR法及流式细胞术检测异补骨脂素对T47D细胞pS2 mRNA及蛋白表达情况的影响。结果1×10-6mol·L-1和1×10-7mol·L-1异补骨脂素能够促进T47D细胞增殖,提高细胞分裂增殖指数,且该作用可被ICI182,780所拮抗;1×10-6mol·L-1和1×10-7mol·L-1异补骨脂素对MDA-MB231细胞增殖无影响。PCR及流式蛋白检测结果显示:1×10-6mol·L-1和1×10-7mol·L-1异补骨脂素可使pS2 mRNA和蛋白表达增加,同时加入ICI182,780可部分拮抗异补骨脂素对pS2表达的诱导和促进作用。结论异补骨脂素具有植物雌激素作用,该效应可通过作用于靶细胞雌激素受体途径实现。

Research on the phytoestrogenic effect and its mechanism of isopsoralen in T47D cells

Aim To study the phytoestrogenic effect and its mechanism of isopsoralen in estrogen receptor (ER) positive T47D and ER negative MDA-MB231 breast cancer cell line.Methods The proliferation rate and cell cycle distribution of T47D and MDA-MB231 cells influenced by isopsoralen were analyzed by MTT and flow cytometry assay respectively.PS2 mRNA level in T47D cells was quantified by Real-time RT-PCR assay.PS2 expression was measured by flow cytometry.Estrogen receptor antagonist ICI180,782 was employed as a tool.Results The proliferation rates and indexes of T47D cells treated with 1×10-6 mol·L-1 and 1×10-7 mol·L-1 isopsoralen were increased.These effects could be blocked by ICI182,780.Meanwhile,isopsoralen didn't show stimulative effects on proliferation rate of MDA-MB231 cells.Real-time RT-PCR and flow cytometry results showed that isopsoralen at 1×10-6 mol·L-1 and 1×10-7 mol·L-1 could induce elevation of pS2 mRNA and protein expression level in T47D cells.These effects could be partly blocked by ICI182,780.Conclusion Isopsoralen has phytoestrogenic effect and its effect is attained mainly via ER pathway.