GPER抑制剂对雌激素长期诱导的MCF-12A乳腺上皮细胞生长和克隆形成的影响
发布时间：2018-08-22 10:48
【摘要】：目的:研究GPER(G protein-coupled estrogen receptor,GPER)抑制剂对雌激素(estrogen,E2)长期作用下乳腺上皮细胞生长和克隆形成的影响,探讨阻断GPER受体防治乳腺癌的可能性。方法:分别使用E2、E2+G15和G15持续作用MCF-12A细胞5周(共传11代)建立细胞模型,通过光镜观察细胞形态的变化、台盼蓝计数法分析细胞生长变化,采用Western blot检测雌激素受体α(estrogen receptorα,ERα)及GPER蛋白表达变化,软琼脂糖克隆形成实验分析细胞克隆形成能力。结果:(1)MCF-12A经E2(10,20和40 nmol·L~(-1))连续处理5周后,其体外生长能力与E2浓度呈一定剂量依赖性,其中,E2(20 nmol·L~(-1))组细胞呈多层重叠样生长,排列紊乱,形态变化最为显著。(2)相比于对照组,E2组中不同代数细胞的ERα蛋白表达明显下调,且呈一定时间依赖性,而GPER蛋白表达未见明显变化。(3)GPER抑制剂G15能抑制E2诱导的细胞生长。(4)G15抑制E2诱导的细胞克隆形成。结论:G15可抑制E2对乳腺上皮细胞MCF-12A的促生长及克隆形成作用,提示GPER抑制剂可能可作为乳腺癌的防治药物。
[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 伪 (ER 伪) 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 伪 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.
【作者单位】： 贵州医科大学天然药物资源优效利用重点实验室;贵州医科大学中药药理学教研室;
【基金】：国家自然科学基金资助项目(编号:81302804) 贵州省科学技术基金(黔科合J字〔2014〕2007号) 贵州省高校优秀科技创新人才支持计划(黔教合KY字〔2015〕492) 2015年度贵州省中医药、民族医药科学技术研究专项课题项目(122) 贵阳市科技计划项目筑科合同[20141001]
【分类号】：R737.9
[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 伪 (ER 伪) 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 伪 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.
【作者单位】： 贵州医科大学天然药物资源优效利用重点实验室;贵州医科大学中药药理学教研室;
【基金】：国家自然科学基金资助项目(编号:81302804) 贵州省科学技术基金(黔科合J字〔2014〕2007号) 贵州省高校优秀科技创新人才支持计划(黔教合KY字〔2015〕492) 2015年度贵州省中医药、民族医药科学技术研究专项课题项目(122) 贵阳市科技计划项目筑科合同[20141001]
【分类号】：R737.9
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