双酚A对子宫肌瘤细胞体外增殖的研究

Effects of bisphenol A on the proliferation of human uterine leiomyoma cells in vitro

目的探讨环境雌激素双酚A(BPA)对人离体子宫肌瘤细胞增殖的影响及相关机制。方法进行人子宫肌瘤细胞原代、传代培养及鉴定。采用噻唑蓝(MTT)法测定用3种剂量的BPA(25×10-7、50×10-7、100×10-7mol/L)分别干预24、48、72h及溶剂对照组细胞的增殖情况;采用反转录-聚合酶链反应(RT-PCR)测定雌激素受体(ER)mRNA,胰岛素样生长因子(IGF)-1mRNA,血管内皮细胞生长因子(VEGF)-1mRNA的表达。结果100×10-7mol/LBPA作用于子宫肌瘤细胞24、48、72h,及50×10-7mol/LBPA作用48、72h可促进子宫肌瘤细胞增殖,并使ER mRNA、IGF mRNA、VEGF mRNA的表达呈上升趋势,与空白对照组比较,差异均有统计学意义(P<0.05,P<0.01)。结论BPA引起细胞增殖可能是通过ER使IGF、VEGF表达增加而引起的,且BPA的促增殖作用存在时间和剂量依赖关系。

Objective To investigate the effects of bisphenol A （BPA）, an environmental estrogen, on proliferation of human uterine leiomyoma cells in vitro and the mechanisms by which it works. Methods The primary culture and subculture of the human uterine leiomyoma cells were performed and identified at each passage by immunocytochemical staining with a monoclonal anti-or-smooth muscle actin antibody. The cells were incubated with BPA （ineluding three dosimetric systems, 25×10^-7 mol/L, 50×10^-7 mol/L and 100×10^-7mol/L） for 24, 48 and 72 h, and the control cells received only the vehicle. The proliferation of cultured human uterine leiomyoma cells was analyzed by MTr assay, and RT-PCR was used to determine the expressions of ER-mRNA, IGF-lmRNA and VEGF-lmRNA. Results Treatment with two doses of BPA （100×10^-7 mol/L for 24, 48 and 72 h; 50×10^-7 mol/L for 48 and 72 h） was shown to stimulate proliferation of human uterine leiomyoma cells and was associated with an increasing trend of ER-mRNA, IGF-mRNA and VEGF-mRNA levels, as compared significantly with the control group （P〈0.05, P〈0.01）. Conclusion Stimulation of human uterine leiomyoma cells by BPA might be attributed to enhanced expression of IGF-1 and VEGF-1 through acting on ER. The proliferation-stimulating effects of BPA displayed a time- and dose- dependent manner.