三种环境雌激素对乳腺癌细胞增殖及凋亡的影响

Influence of Environmental Estrogens on Cell Proliferation and Apoptosis in T47D Cells

[目的 ]探讨环境中常见的三种类雌激素壬基酚 (NP)、双酚A(BisA)和邻苯二甲酸二丁酯 (DBP)影响人乳腺癌细胞T47D增殖和凋亡的分子生物学作用机制。 [方法 ]T47D细胞在含 10 %胎牛血清的DMEM培养液中进行常规传代培养 ,实验前将细胞转移至无酚红DMEM(含 5 %活性碳葡聚糖苷处理过的FBS)中继续培养 5d ,收集细胞 ,PBS洗涤后接种于内置盖玻片的 6孔板或 75ml培养瓶中 ,用 3 2× 10 -7mol/LNP、3 2× 10 -7mol/LBisA及 3 2× 10 -6mol/LDBP对细胞分别处理 72h ,用半定量RT PCR技术观察这 3种化合物对核增殖抗原PCNA、bcl 2及baxmRNA表达的影响 ,并用免疫组化方法对结果进行验证。 [结果 ]与溶剂对照组相比 ,NP和BisA对T47D细胞处理 72h可显著抑制baxmRNA表达而促进bcl 2和PCNAmRNA的表达 ,随后的免疫组化实验也证实了这一结果 ;DBP可促进PCNAmRNA及蛋白的表达 ,但对bcl 2及bax的表达未见统计学意义。 [结论 ]环境雌激素NP和BisA可通过调节PCNA、bcl 2及bax的表达而调节乳腺癌T47D细胞的增殖和凋亡作用。

Objective To investigate the breast cancer T47D cell proliferation and apoptosis induced by environmental estrogens (n-4-noniphenol,NP; bisphenol A,BisA; dibutylphthalate,DBP).[Methods] Cells were maintained in DMEM medium with 10% fetal bovine serum. Five days before the beginning of experiments,the cells were seeded in phenol red-free DMEM medium containing 5% charcoal dextran-treated FBS. The cells were harvested,and seeded in 6-well culture plates or in 75 ml flacks. After NP,BisA and DBP treatments for 72 h,the cells were harvested,and mRNA and protein expression of PCNA,bcl-2 and bax were detected by reverse transcription-polymerase chain reaction and immunohistochemistry,respectively. [Results] Compared with the vehicle control,at the concentration of 32×10 -7mol/L,NP and BisA could significantly up-regulate bcl-2 and PCNA mRNA expression and down-regulate bax mRNA expression,and 32×10 -6 mol/L DBP had no effects on mRNA expression of bax and bcl-2,but could stimulated PCNA mRNA expression. These results were further confirmed by following immunohistochemistry. [Conclusion] PCNA,bcl-2 and bax pathway might involve in cell proliferation and apoptosis events regulated by environmental estrogens in breast cancer T47D cells.