摘要
目的 探讨G蛋白耦联雌激素受体(GPER)对正常前列腺上皮细胞增殖及自噬的影响。 方法 利用CCK8,观察雌二醇、G1对两种前列腺上皮细胞BPH-1及RWPE-1增殖的影响,通过Western印迹、CYTO-ID自噬检测试剂检测加入雌二醇、G1及同时加入G1及G15对前列腺上皮细胞的自噬活动的作用。 结果 加入雌二醇、G1后24 h开始,雌二醇组及G1组的吸光度(A)值均低于空白对照组(P<0.01);显示雌二醇及G1对BPH-1及RWPE-1细胞处理有抑制作用,且随时间延长,抑制效应增加。雌二醇组及G1组中的LC3-Ⅱ/LC3-Ⅰ比值及CytoID绿色荧光强度均较空白对照组低(P<0.01),G1+G15组较G1组的LC3-Ⅱ/LC3-Ⅰ比值及CytoID绿色荧光强度高(P<0.05)。 结论 GPER的激活可抑制细胞的自噬及增殖。
Objective To investigate the effect of G protein-coupled estrogen receptor (GPER) on proliferation and autophagy in prostatic epithelial cells. Methods Cell Counting Kit-8 (CCK8) assay was used to observe the growth of BPH-1 and RWPE-1 cells after treating with or not with estradiol or GPER selective agonist G1. Autophagy was quantified with Western blot and Cyto-ID autophagy detection kit after treating with estradiol, G1 or both G1 and G15 in the same cells. Results The OD value in estrogen group and G1 group was significantly lower than those in control group (P<0.01). Compared with control group, Western blot and Cyto-ID green reagent staining revealed that the ratio of LC3Ⅱ/LC3Ⅰ and the relative fluorescence intensity of BPH-1 and RWPE-1 cells were decreased in G1 group and estradiol group (P<0.01). Pretreatment with G15 reversed the effect of G1 (P<0.05). Conclusion The activation of GPER leads to the inhibition of autophagy and cell proliferation.
作者
董卫民
卢剑铭
朱建国
叶剑恒
冯源发
何慧婵
Dong Weimin;Lu Jianming;Zhu Jianguo;Ye Jianheng;Feng Yuanfa;He Huichan(Department of Urology,the First Affiliated Hospital of Guangzhou Medical College,Guangdong Key Laboratory of Clinical Molecular Medicine and Molecular Diagnosis,Guangzhou 510180,China;Department of Urology,Guizhou Provincial People's Hospital,Guizhou 550002,China;Department of Urology,the First Affiliated Hospital of Guangzhou Medical College,Guangdong Key Laboratory of Urology,Guangzhou 510230,China)
出处
《中华医学杂志》
CAS
CSCD
北大核心
2019年第6期419-422,共4页
National Medical Journal of China
基金
国家自然科学基金(81470983,81360119)
广东省科技计划项目(2013B021800055)
广东省临床分子医学及分子诊断实验室重点实验室项目、广东省泌尿外科重点实验室项目(2010A060801016)
广州市科技计划项目(2014J4100072).
