GPR30介导双酚A促进小鼠GC-1细胞增殖  

GC-1 cell proliferation promoted by bisphenol A via G protein-coupled receptor 30

作　　者：陈远彬[1] 黄卫人[2] 秦达念[1] 

机构地区：[1]汕头大学医学院,汕头515000 [2]北京大学深圳医院男性生殖与遗传广东省重点实验室

出　　处：《中国男科学杂志》2012年第4期3-8,共6页Chinese Journal of Andrology

摘　　要：目的研究环境雌激素双酚A（bisphenol A，BPA）通过G蛋白偶联受体30（Gprotein-coupled receptor 30，GPR30）对小鼠精原细胞系GC-1细胞的影响及发生机制。方法（1）对GC-1细胞分组（分为对照组，BPA＋ICI，BPA＋ICI＋GPR30 siRNA，BPA＋ICI＋AG1478，BPA＋ICI＋PD98059，G-1六个组）进行处理，培养0h，24h，48h，72h，96h后采用MTT比色法检测细胞活力。（2）分别用Gpr30 siRNA沉默Gpr30基因，AG1478阻断EGFR（epidermal growth factor receptor），PD98059阻断ERK（extracellular signal-regulated kinases），然后BPA处理GC-1细胞，5min后Western blot检测ERK-1／-2磷酸化水平和Real-time PCR检测转录因子c-Fos变化程度，48h后检测周期蛋白基因Cyclin D1表达情况。结果与对照组相比，BPA处理组促进GC-1细胞增殖，c-Fos和Cyclin D1基因表达上升（P〈0．05）。细胞在上述三个水平阻断处理后，BPA不引起相应表达差异。结论在小鼠GC-1细胞，BPA诱导GPR30转活EGFR，通过激活MAPK／ERK—c—Fos信号调节，上调周期蛋白基因Cyclin D1表达，促进细胞增殖。Objective To investigate the effect and mechanisms induced by BPA via GPR30 in GC-1 cells. Methods 1. GC-1 cells were divided into 6 groups and pretreated with ICI182780, then the cells were prior to being silenced GPR30, or pre-incubated with AG1478 or PD98059 respectively followed by BPA treated for 0-96 h. The cell viability was detected using MTT assays. The control group was added with DMSO in which the final concentration was 0.1%, and the group added with G-1 was the positive control group. 2. The cells were treated as above, and the phosphorylation of ERK-1/-2 was examined using Western blot analysis and the expression level of c-Fos mRNA using Real-time PCR analysis after BPA treated for 5 min, besides, the expression level of Cyclin D1 mRNA was detected using Real-time PCR analysis after BPA treated for 48 h. Results BPA promoted GC-1 cell proliferation compared with the control group and increased the expression level of c-Fos and Cyclin D1 genes, P〈0.05. Prior to silenced GPR30 or pretreated with AG1478 or PD98059, the cell proliferation induced by BPA was blocked. Conclusion The complex of BPA bound to GPR30 transactivated EGFR and activated MAPK-ERK-c-Fos pathway in the GC-1 cells, and increased the expression of Cyclin D 1 mRNA, resulted in cell proliferation.

关 键 词：双酚A 受体 G-蛋白偶联 GC-1细胞 MAP激酶信号系统 

分 类 号：R361.3[医药卫生—病理学] R321.1[医药卫生—基础医学]