阿片类生长因子受体抑制雄激素依赖性前列腺癌LNCaP细胞的生长

Opioid growth factor receptor depresses growth of androgen-dependent prostate cancer LNCaP cells

目的:观察阿片类生长因子受体(opioid growth factor receptor,OGFr)对雄激素依赖性前列腺癌LNCaP细胞活力的影响。方法:将LNCaP细胞随机分为非转染组、空白质粒组(转染pcDNA3.1空白质粒)和OGFr质粒组(转染表达质粒pcDNA3.1-OGFr),RT-qPCR和Western blot检测转染效率;10 nmol/L二氢睾酮(dihydrotestosterone,DHT)分别处理LNCaP细胞24、48、72和96 h后,CCK-8法检测细胞活力,流式细胞术检测细胞周期及凋亡的变化,RT-qPCR和Western blot检测细胞周期调控蛋白——细胞周期蛋白依赖性激酶2(cyclin-dependent kinase 2,CDK2)、细胞周期蛋白E(cyclin E)和p21的表达。结果:相对于非转染组,pcDNA3.1-OGFr转染显著增加LNCaP细胞OGFr的mRNA和蛋白表达(P<0.05);10 nmol/L DHT呈时间依赖性促进LNCaP细胞的活力(P<0.05),而DHT对OGFr质粒组细胞活力的增强效应显著降低(P<0.05);在DHT的作用下,OGFr质粒组细胞发生G_(0)/G_(1)期阻滞(P<0.05),但细胞凋亡率无显著差异;CDK2表达下调(P<0.05),p21表达上调(P<0.05),而cyclin E表达无显著差异。结论:OGFr诱导的细胞周期阻滞可抑制雄激素DHT对人前列腺癌LNCaP细胞的生长效应。

AIM:To investigate the effect of opioid growth factor receptor(OGFr)on the growth of androgendependent human prostate cancer LNCaP cells.METHODS:Cultured LNCaP cells were randomly divided into nontransfection group,pcDNA3.1 empty plasmid group and pcDNA3.1-OGFr group.Transfection efficiency was evaluated by RTqPCR and Western blot analysis.The viability of LNCaP cells was measured by CCK-8 assay in the presence of 10 nmol/L dihydrotestosterone(DHT)at different time points(24,48,72 and 96 h).Cell cycle and apoptosis were examined by flow cytometry,and the expression of cyclin-dependent kinase 2(CDK2),cyclin E and p21 at mRNA and protein levels were determined by RT-qPCR and Western blot.RESULTS:The mRNA and protein levels of OGFr exhibited a marked increase in pcDNA3.1-OGFr group compared with nontransfection group(P<0.05).Treatment with 10 nmol/L DHT promoted the viability of LNCaP cells in a time-dependent manner(P<0.05).However,the effect of DHT on LNCaP cell viability was partly abolished in pcDNA3.1-OGFr group(P<0.05).The LNCaP cells with OGFr over-expression exhibited significant blockage in G_(0)/G_(1) phase in the presence of 10 nmol/L DHT(P<0.05),but no effect on apoptosis.Furthermore,the expression of CDK2 at mRNA and protein levels in pcDNA3.1-OGFr group was decreased(P<0.05),p21 expression was increased(P<0.05),and no detectable expression change in cyclin E was found.CONCLUSION:OGFrinduced cell cycle arrest has a potent inhibitory effect on DHT-induced growth of human prostate cancer LNCaP cells.