中文摘要：

目的研究六氟双酚A（bisphenol AF,BPAF）诱导T47D乳腺癌细胞中细胞外调节蛋白激酶（extracellular regulated protein kinases,ERK）激活的分子机制,探讨雌激素受体α（estrogen receptorα,ERα）在BPAF诱导ERK激活过程中的作用。方法通过蛋白免疫印迹实验（Western Blot）检测ERK磷酸化水平,研究BPAF对ERK磷酸化的诱导作用。运用表达ERαshRNA的慢病毒感染细胞,下调表达ERα,研究ERα在BPAF激活ERK过程中的作用。结果Western Blot结果显示,BPAF在浓度为50 nmol/L时即可诱导ERK磷酸化水平显著升高（P〈0.05）,并且随着剂量的升高呈现出剂量-效应关系。当BPAF浓度达到1μmol/L时ERK磷酸化水平达到最高（P〈0.05）。BPAF诱导ERK激活时间效应实验中,BPAF处理细胞5~60 min时,ERK磷酸化水平均显著升高（P〈0.05）,在BPAF处理细胞15 min时,ERK磷酸化水平达到最高。在运用表达ERαshRNA的慢病毒感染细胞后,ERα的表达水平显著降低（P〈0.05）。与正常细胞相比,在下调表达ERα的细胞中,BPAF对ERK磷酸化的诱导作用显著降低（P〈0.05）。结论 BPAF能够诱导ERK磷酸化,并且呈现剂量-效应关系。ERα在BPAF诱导的ERK激活过程中起到重要作用。

英文摘要：

Objective To evaluate the effect of bisphenol AF（ BPAF） on phosphorylation of extracellular regulated protein kinases（ ERK） in T47 D human breast cancer cells,and explore the role of estrogen receptor α（ ERα） in BPAF-induced ERK activation.Methods ERK phosphorylation was evaluated by Western Blot. Lentivirus-mediated shRNA expression were utilized to knockdown ERα.Results BPAF significantly stimulated ERK phosphorylation in a dose-response manner at the dose range from 50 nmol / L to 10 μmol / L（ P〈0. 05） of BPAF. 1 μmol/L significantly stimulated ERK phosphorylation during 5 ~ 60 min（ P〈0. 05）,and showed strongest induction at 15 min. Silencing of ERα by shRNA substantially decreased BPAF-induced ERK phosphorylation compared to scramble shRNA（ P〈0. 05）. Conclusion According to the dose and time response studies,BPAF could significantly stimulate ERK phosphorylation in T47 D human breast cancer cells. ERα played an important role on BPAF-induced ERK phosphorylation,indicating the endocrine disrupting effect of BPAF might be mediated through activation of ERK signaling.