Liu Huidi, Yan Yan, Wen Haixia, Jiang Xueli, Cao Xuefeng, Zhang Guangmei, Liu Guoyi
Laboratory of Reproductive Endocrinology, Department of Physiology, Harbin Medical University, Harbin, 150081, China.
Cell Biol Int. 2014 May;38(5):631-8. doi: 10.1002/cbin.10243. Epub 2014 Jan 30.
G protein-coupled estrogen receptor (GPER) is recently identified as a membrane-associated estrogen receptor that mediates non-genomic effects of estrogen. Our previous immunohistochemistry study found an association between GPER and the proliferation of epithelial ovarian cancer. However, the contributions and mechanisms of GPER in the proliferation of ovarian cancers are not clear. We have examined the role of GPER in estrogen receptor α (ERα)-negative/GPER positive OVCAR5 ovarian cancer cell line. MTT assay was used to detect cell proliferation. BrdU incorporation assay was used to measure the cells in S-phase. Protein expression of marker genes of proliferation, cell cycle and apoptosis were examined by Western blot. The results showed that 17β-estradiol and selective GPER agonist G-1 stimulated the proliferation of OVCAR5 cells and increased the cells in S-phase. Both ligands upregulated the protein levels of c-fos and cyclin D1. Small interfering RNA targeting GPER or G protein inhibitor pertussin toxin (PTX) inhibited basal cell proliferation and attenuated 17β-estradiol- or G-1-induced cell proliferation. GPER mediated cell growth was also associated with the apoptosis of OVCAR5 cells. These findings suggest that GPER has an important function in the proliferation of ovarian cancer cells lacking ERα. GPER might be a promising therapeutic target in ovarian cancer.
G蛋白偶联雌激素受体（GPER）最近被鉴定为一种膜相关雌激素受体，介导雌激素的非基因组效应。我们之前的免疫组织化学研究发现GPER与上皮性卵巢癌的增殖之间存在关联。然而，GPER在卵巢癌增殖中的作用和机制尚不清楚。我们研究了GPER在雌激素受体α（ERα）阴性/GPER阳性的OVCAR5卵巢癌细胞系中的作用。采用MTT法检测细胞增殖。采用BrdU掺入法检测S期细胞。通过蛋白质印迹法检测增殖、细胞周期和凋亡标记基因的蛋白表达。结果显示，17β-雌二醇和选择性GPER激动剂G-1刺激OVCAR5细胞增殖并增加S期细胞数量。两种配体均上调c-fos和细胞周期蛋白D1的蛋白水平。靶向GPER的小干扰RNA或G蛋白抑制剂百日咳毒素（PTX）抑制基础细胞增殖，并减弱17β-雌二醇或G-1诱导的细胞增殖。GPER介导的细胞生长也与OVCAR5细胞的凋亡有关。这些发现表明，GPER在缺乏ERα的卵巢癌细胞增殖中具有重要作用。GPER可能是卵巢癌一个有前景的治疗靶点。
