發(fā)布時間：2018-08-22 10:48

【摘要】：目的:研究GPER(G protein-coupled estrogen receptor,GPER)抑制劑對雌激素(estrogen,E2)長期作用下乳腺上皮細胞生長和克隆形成的影響,探討阻斷GPER受體防治乳腺癌的可能性。方法:分別使用E2、E2+G15和G15持續(xù)作用MCF-12A細胞5周(共傳11代)建立細胞模型,通過光鏡觀察細胞形態(tài)的變化、臺盼藍計數(shù)法分析細胞生長變化,采用Western blot檢測雌激素受體α(estrogen receptorα,ERα)及GPER蛋白表達變化,軟瓊脂糖克隆形成實驗分析細胞克隆形成能力。結(jié)果:(1)MCF-12A經(jīng)E2(10,20和40 nmol·L~(-1))連續(xù)處理5周后,其體外生長能力與E2濃度呈一定劑量依賴性,其中,E2(20 nmol·L~(-1))組細胞呈多層重疊樣生長,排列紊亂,形態(tài)變化最為顯著。(2)相比于對照組,E2組中不同代數(shù)細胞的ERα蛋白表達明顯下調(diào),且呈一定時間依賴性,而GPER蛋白表達未見明顯變化。(3)GPER抑制劑G15能抑制E2誘導(dǎo)的細胞生長。(4)G15抑制E2誘導(dǎo)的細胞克隆形成。結(jié)論:G15可抑制E2對乳腺上皮細胞MCF-12A的促生長及克隆形成作用,提示GPER抑制劑可能可作為乳腺癌的防治藥物。
[Abstract]:Aim: to study the effects of GPER (G protein-coupled estrogen receptor inhibitor on the growth and clone formation of breast epithelial cells under the long-term action of estrogen E2, and to explore the possibility of blocking GPER receptor in the prevention and treatment of breast cancer. Methods: the cell model of MCF-12A cells was established by continuous treatment of E2G15 and G15 for 5 weeks (11 passages). The changes of cell morphology were observed by light microscope, and the changes of cell growth were analyzed by trypan blue counting. The expression of estrogen receptor 偽 (ER 偽) and GPER protein were detected by Western blot, and the ability of cell clone formation was analyzed by soft agarose clone forming assay. Results: (1) after MCF-12A was treated with E2 (10 ~ 20 and 40 nmol L ~ (-1) for 5 weeks, there was a dose-dependent relationship between in vitro growth ability and E _ 2 concentration. The cells in E _ 2 (20 nmol L ~ (-1) group grew in a multilayer overlapping manner, and the arrangement was disordered. (2) compared with the control group, the expression of ER 偽 protein in different algebraic cells was down-regulated in a time-dependent manner. The expression of GPER protein did not change significantly. (3) GPER inhibitor G15 inhibited E2-induced cell growth and (4) G15 inhibited E2-induced cell clone formation. Conclusion the growth and clone formation of MCF-12A in breast epithelial cells can be inhibited by E _ 2, suggesting that the GPER inhibitor may be used as a preventive and therapeutic drug for breast cancer.
【作者單位】： 貴州醫(yī)科大學(xué)天然藥物資源優(yōu)效利用重點實驗室;貴州醫(yī)科大學(xué)中藥藥理學(xué)教研室;
【基金】：國家自然科學(xué)基金資助項目(編號:81302804) 貴州省科學(xué)技術(shù)基金(黔科合J字〔2014〕2007號) 貴州省高校優(yōu)秀科技創(chuàng)新人才支持計劃(黔教合KY字〔2015〕492) 2015年度貴州省中醫(yī)藥、民族醫(yī)藥科學(xué)技術(shù)研究專項課題項目(122) 貴陽市科技計劃項目筑科合同[20141001]
【分類號】：R737.9

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