GPR30在子宫平滑肌瘤细胞增殖中的作用研究  

Expression of GPR30 in leiomyoma cells of uterus and its relation to cell proliferation

作　　者：徐学麟[1] 林俊[2] 江秀秀[2] 杨艳峰[2] 詹宏[2] 李敏[2] 

机构地区：[1]宁波大学医学院附属医院妇产科,浙江宁波315000 [2]浙江大学医学院附属妇产科医院妇产科,浙江杭州310006

出　　处：《实用肿瘤杂志》2014年第3期234-238,共5页Journal of Practical Oncology

摘　　要：目的探讨跨膜G蛋白偶联受体30(G protein-coupled receptor30,GPR30)在子宫平滑肌瘤细胞中的表达及其对肌瘤中平滑肌细胞的增殖作用。方法培养子宫平滑肌瘤原代细胞,应用免疫组织化学、免疫荧光及蛋白质印迹法检测GPR30的表达。应用蛋白印迹检测GPR30干扰后,其下游信号通路的变化。应用MTT检测在雌激素刺激下,正常子宫平滑肌细胞及平滑肌瘤细胞的生长活性。结果子宫平滑肌瘤细胞中有GPR30蛋白的高表达,其主要位于细胞质中,在细胞核上无明显表达。在未处理组平滑肌细胞中,雌激素刺激雌激素信号通路中的ERK1/2表达增强。在GPR30干扰后的平滑肌细胞中,雌激素刺激后ERK1/2表达无显著改变。MTT结果表明在雌激素作用下,未处理组细胞存活和生长率显著增加;干扰组细胞的存活和生长率显著下降。结论 GPR30在子宫肌瘤平滑肌细胞的细胞质中高表达,可能参与子宫平滑肌细胞的增殖。Objective To investigate the expression of G protein-coupled receptor 30 （GPR30） in leiomyoma cells of uterus and its relation to cell proliferation. Methods Myometrial biopsy samples were obtained from women with leiomyomas, and leiomyoma cells were primarily cultured. Immunohistochemistry, immunofluorescence and Western blot were used to detect GPR30 expression in uterine leiomyomas and normal myometrial tissues. The downstream signaling pathways after the interference of GPR30 were detected with Western blot. The proliferation of myometrial cells and leiomyoblastoma cells after the treatment of estrogen was measured by MTT. Results GPR30 protein was expressed highly in leiomyomas cells and mainly localized in cytoplasm, but not in the nucleus. The GPR30 protein level in uterine leiomyomas was significantly higher than that in normal myometrium （ P 〈 0.05 ）. E2 stimulated rapid extranuclear signaling in myometrial explants was observed, with increased extracellular sigual-regulated kinase （ERK1/2） phosphorylation. This effect was inhibited by pretreatment with GPR30 siRNA. MTT showed increased cell growth and survival in untreated cells, but opposite effect was observed in cells pretreated with GPR30 siRNA. Conclusion GPR30 is highly expressed in the cytoplasm of leiomyomas cells and may be involved in the proliferation of human myometrium cells.

关 键 词：平滑肌瘤 病理学 受体 G-蛋白偶联 雌激素受体 药理学 子宫 ERK-1 2 细胞增殖 药物作用 氮蓝四唑 药代动力学 比色法 

分 类 号：R737.33[医药卫生—肿瘤]