G蛋白耦联受体C家族6组A亚型在前列腺癌细胞增殖和凋亡中的作用  

The role of GPRC6A in prostate cancer cell on proliferation and apoptosis

作　　者：孙心旖 盛彬[1] 陈瑶[1] 李莉[1] 杨建一[1] 杜圣家[1] 刘铭[1] SUN Xin-yi;SHENG Bin;CHEN Yao;LI Li;YANG Jian-yi;DU Sheng-jia;LIU Ming(School of Basic Medical Science,Shanxi Medical University,Taiyuan 030001,Shanxi,China)

机构地区：[1]山西医科大学基础医学院,太原 030001

出　　处：《医学研究生学报》2019年第2期119-125,共7页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81302220);山西省青年科学基金(2014021037?1)

摘　　要：目的前列腺癌(PCa)在我国的发病率逐年上升。G蛋白耦联受体C家族6组A亚型(GPRC6A)是近年来发现的PCa易感基因。文章探讨GPRC6A及其介导的细胞外信号调节蛋白激酶(ERK)在前列腺癌LncapC4?2细胞增殖与凋亡中的作用。方法实验分为LncapC4?2 PCDH组(仅转染空载体pCDH)、LncapC4?2 GPRC6A组(仅转染pCDH?GPRC6A)和LncapC4?2 GPRC6A+U0126组(转染pCDH?GPRC6A并加入U0126处理),通过CCK8检测细胞增殖变化,流式细胞仪检测细胞周期和凋亡改变以及Western blot方法检测细胞周期与凋亡相关蛋白的变化。结果 Western blot法结果表明,与LncapC4?2 PCDH组比较,LncapC4?2 GPRC6A组的GPRC6A和P?ERK表达增加(P<0.05);与LncapC4?2 GPRC6A组比较,LncapC4?2 GPRC6A+U0126组的P?ERK的表达明显降低(P<0.05)。CCK8检测表明LncapC4?2 GPRC6A组相较LncapC4?2 PCDH组增殖加快(P<0.05),而LncapC4?2 GPRC6A+U0126组相较LncapC4?2 GPRC6A组增殖明显减慢(P<0.05)。流式细胞仪检测结果显示LncapC4?2 GPRC6A组的G1/(S+G2)值较LncapC4?2 PCDH组明显降低(P<0.05);LncapC4?2 GPRC6A+U0126组的G1/(S+G2)值较LncapC4?2 GPRC6A组明显升高(P<0.05)。Western blot结果中LncapC4?2 GPRC6A组的CyclinD1蛋白表达(0.88±0.04)较LncapC4?2 PCDH组(0.66±0.02)明显升高(P<0.05),而LncapC4?2 GPRC6A+U0126组的CyclinD1蛋白表达(0.71±0.02)较LncapC4?2 GPRC6A组明显降低(P<0.05)。流式细胞仪检测凋亡结果表明,LncapC4?2 GPRC6A组的凋亡比例(3.64%)较LncapC4?2 PCDH组(9.00%)和LncapC4?2 GPRC6A+U0126组(19.57%)明显降低(P<0.05)。同时Western blot检测结果表明,LncapC4?2 GPRC6A组的Bcl2的表达比LncapC4?2 PCDH组高,活化Caspase3的表达则降低(P<0.05);LncapC4?2 GPRC6A+U0126组相对于LncapC4?2 GPRC6A组Bcl2的表达降低,活化Caspase3的表达则增高(P<0.05)。结论 GPRC6A可能通过ERK信号通路促进PCa细胞增殖,抑制细胞凋亡,而参与PCa的发展过程。Objective The incidence of prostate cancer(PCa)is increasing year by year in China.G protein coupled receptor family C,group 6,member A(GPRC6A)is a susceptible gene of PCa found in recent years.To investigate therole of GPRC6A and extracellular signal-regulated kinase(ERK)mediated by GPRC6A in the proliferation and apoptosis of PCaLncapC4-2 cells,and provide new ideas for targeted therapy of PCa.Methods The experiment was divided into the LncapC4-2 PCDH group(only the empty vector pCDH was transfected),the LncapC4-2 GPRC6A group(only the pCDH-GPRC6A was transfected)and the LncapC4-2 GPRC6A+U0126 group(pCDH-GPRC6A was transfected and treated with U0126).The proliferation of LncapC4-2 cells was detected by using CCK8 kit.The changes of cell cycle and apoptosis were detected using flow cytometry.Cell cycle and apoptosis-related proteins were evaluated by Western blot.Results The result of Western blots showed the expression of GPRC6A and P-ERK in LncapC4-2 GPRC6A group washigher than that in LncapC4-2 PCDH group(P<0.05),and the expression of P-ERKin LncapC4-2 GPRC6A+U0126 group was evidently lower than that in LncapC4-2 GPRC6A group(P<0.05).The result of CCK8 tests showed the cell proliferation ability in LncapC4-2 GPRC6A group was higher than that in LncapC4-2 PCDH group,and the proliferation ability in LncapC4-2 GPRC6A+U0126 group was lower than that in LncapC4-2 GPRC6A group,and there was statistical difference between two groups(P<0.05).Flow cytometry results showed the ratio of G1/(G2+S)in LncapC4-2 GPRC6A groupwas evidently lower than that in LncapC4-2 PCDH group(P<0.05),and the ratio of G1(/G2+S)in LncapC4-2 GPRC6A+U0126 group was evidently higher than that in LncapC4-2 GPRC6A group(P<0.05).Western blot results showed that the expression of CyclinD1 were evidently increased in LncapC4-2 GPRC6A group(0.88±0.04)compared to LncapC4-2 PCDH group(0.66±0.02,P<0.05),the expression CyclinD1 wasevidently reduced in LncapC4-2 GPRC6A+U0126 group(0.71±0.02)compared with LncapC4-2 GPRC6A group(P<0.05).The ratio of apop

关 键 词：G蛋白耦联受体C家族6组A亚型 前列腺癌 细胞外信号调节蛋白激酶 

分 类 号：R737.25[医药卫生—肿瘤]