松花江水中有机提取物对MCF-7细胞增殖、迁移及对Bcl-2/Bax蛋白表达的影响

目的： 探讨松花江水源水中有机提取物的环境雌激素效应及其诱导人乳腺癌MCF-7细胞增殖的作用机制。方法：于2011年3月采集松花江上游四方台水源水样1 000 L，用GDX-102大孔树脂提取水中有机物，分别以相当于原水量25、100、400、1 600和3 200 mL的水中有机提取物与MCF-7细胞共培养，通过MTT实验检测细胞的增殖情况，划痕损伤实验和transwell迁移实验观察MCF-7细胞侵袭迁移，并采用免疫细胞化学法检测有机提取物对Bcl-2和Bax蛋白表达的影响。结果：与阴性对照组比较，随着有机提取物浓度升高雌激素效应逐渐增强，差异均有统计学意义 (P<0.05)。当有机提取物含量相当于原水量1 600 mL时雌激素效应最强，促进细胞侵袭迁移作用亦最强。Bcl-2和Bax蛋白在MCF-7细胞中均表达，与阴性对照组比较，随着有机提取物浓度升高Bcl-2蛋白表达率逐渐增高 (P<0.05)，而Bax蛋白表达率逐渐降低 (P<0.05)。结论：松花江上游四方台水源水中有机提取物可促进MCF-7细胞增殖和侵袭迁移，且可能通过调控Bcl-2/Bax蛋白比例促进MCF-7细胞增殖。

The influence of organic extracts in Songhua River on proliferation and migration of MCF-7 cell as well as expression of Bcl-2/Bax protein

OBJECTIVE: Exploring the environmental estrogenic effect and mechanism of cell proliferation induced by organic extracts from Songhua River. METHODS：1 000 L water sample was collected from upstream water (Sifangtai source water)of Songhua River in March 2011. Organic substances were extracted by GDX-102 resin. MCF-7 cells were cultured with different concentrations of organics extracts (the concentrations were equivalent to those from 25，100，400，1 600 and 3 200 mL of river water). We measured estrogenic effect by MTT assay and investigated invasion and migration of MCF-7 cells by wound healing assay and transwell assay. We also explored the influence of organic extracts on Bcl-2 and Bax protein expressions by immunocytochemical method. RESULTS：The estrogenic effect was gradually enhanced with increasing concentration of organic extracts，significantly different compared to negative control group (P<0.05). The estrogenic effect and promotion of cell invasion and migration were the strongest when the content of organic extracts was equivalent to 1 600 mL of river water. Both Bcl-2 and Bax proteins could be expressed in MCF-7 cells. Comparing to negative control，with increasing concentration of organic extracts， the expression of Bcl-2 protein was gradually increased while that of Bax protein decreased. CONCLUSION：Organic extracts from upstream water(Sifangtai source water) of Songhua River could promote the proliferation of MCF-7 cells by regulating the ratio of Bcl-2 and Bax proteins，and stimulate invasion and migration of MCF-7 cells.