目的:观察G蛋白偶联雌激素受体(GPER)特异性抑制剂G15对17β-雌二醇(E2)剌激转化细胞增殖能力的影响及其机制初探.方法:自制乳腺癌细胞,乳腺癌MCF-10A细胞用50nmol/LE2连续刺激至第13代,以DMSO刺激作为对照(均标记为P13);显微镜观察细胞转化形态,将DMSO及E2处理后的P13分别接种到裸鼠皮下,接种后1周观察接种部位的成瘤情况;将自制乳腺癌细胞分为E2组和10 μmol/L G15组,以MCF-7细胞作为阳性对照细胞,用平板克隆形成实验观察细胞的增殖能力.结果:镜下可见P13代MCF-H10A细胞接触抑制消失、细胞胞体变大、出现克隆小球,裸鼠乳腺脂肪垫皮下成瘤(7/10),对照无瘤形成;平板克隆实验中,自制乳腺癌细胞、MCF-7细胞与各自阴性对照比较,E2刺激组的增殖加快,差异有统计学意义(P<0.05);与各自E2刺激组比较,G15抑制组增殖能力下降,差异有统计学意义(P<0.05).结论:GPER介导E2刺激转化细胞增殖能力增加过程.%Objective: To assess the role of inhibitor G15 in G protein-coupled estrogen receptor (GPER) in malignantly transformed mammary epithelial cells. Methods: Non-tumorigenic MCF-10A mammary epithelial cells were used as the research models. MCF-7 breast cancer cells were used as the positive control in in vitro assay. Model cells were cultured up to the thirteenth passage, marked as P13. The morphology of treated cells was observed under the microscope, and cell proliferation was detected by plate colony formation assay. Then tumorigenicity was detected in nude mice. Results: In cells treated with E2 (50nM/L), cell contact inhibition disappeared and cell bodies became larger and fusiform. Transformed cells and MCF-7 cells were further stimulated with E2(with DMSO as controls) and cell proliferation greatly increased(P＜0.01) and this effect could be dramatically suppressed by G15. E2-treated model cells were tumorigenic (7/10) significantly in nude mice (with controls as 0/5). Conclusion: Novel estrogen receptor GPER may be led to promote the proliferation ability of E2-induced transformed cells.
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