BPA激活GPER-EGFR-ERK1/2信号通路诱导乳腺癌细胞增殖

Involvement of GPER-EGFR-ERK1/2 in Bisphenol A Induced Breast Cancer Cell Proliferation

目的：研究雌激素G蛋白偶联受体（G protein-coupled estrogen receptor,GPER）-表皮生长因子受体（epidermal growthfactor receptor,EGFR）-细胞外调节蛋白激酶1/2（extracellular regulated protein kinases,ERK）信号通路在双酚A促乳腺癌细胞SKBR-3增殖中的作用。方法：采用CCK8试剂盒检测SKBR-3细胞的增殖情况,利用Western Blot观察ERK1/2磷酸化变化。结果：与对照组相比,10-11~10-7 M浓度的双酚A具有促乳腺癌细胞增殖作用,10-9M浓度的双酚A可诱导细胞增殖最大效应（细胞活力高于对照组约38.84%,P〈0.001）;预孵GPER、EGFR、ERK1/2特异性抑制剂G15、AG-1478、PD98059后,双酚A诱导的乳腺癌细胞增殖明显减少,细胞活力与单纯双酚A（10-9 M）处理相比降低约14.27%、12.23%和17.98%（P〈0.05）;双酚A（10-9 M）处理细胞0.5、1、3小时后,发现磷酸化的细胞外信号调节激酶（p-ERK）的表达明显高于对照组,双酚A可快速激活ERK1/2;而阻断GPR30和EGFR后,ERK1/2的磷酸化表达减少。结论：双酚A诱导的乳腺癌细胞增殖可能与GPR30-EGFR-ERK1/2信号通路有关,但不是唯一通路。深入研究双酚A对促进乳腺癌细胞增殖机制,可能为乳腺癌的防治提供新的方向。

Objective： To explore the effect of Bisphenol A on the proliferation of human breast cancer cells（SKBR-3） via GPER-EGFR-ERK1/2 signaling pathway. Methods： CCK8 assay was used to detect the proliferation of SKBR-3 cells, and Western Blotting was applied to observe phosphorylation of ERK1/2. Results： Cell proliferation increased significantly with treatment of 10-11-10-7M BPA after 24 h, and 10-9M BPA can induce cell proliferation maximum effect（cell viability higher approximately 38.84 % compared with the control, P〈0.001）; with pre-incubation of G15, AG-1478 and PD98059（specific antagonist of GPER, EGFR and ERK1/2）, cell proliferation decreased remarkably. Cell viability had a decrease of approximately 14.27 %, 12.23 % and 17.98 % compared with BPA（10-9M） treatment（P〈0.05）; 0.5, 1, 3 h after BPA（10-9M） treatment, phosphorylated extracellular signal-regulated kinase（p-ERK）expression increased significantly,indicating that ERK 1/2 was activated rapidly; after blocking GPR30 and EGFR, ERK 1/2phosphorylation decreased. Conclusions： GPR30-EGFR-ERK 1/2 signaling pathway was involved in BPA-induced breast cancer cell proliferation. In-depth research of BPA to breast cancer cell proliferation may provide a new direction for the prevention and treatment of breast cancer.