雄激素受体调控的lncRNA ARLNC1对前列腺癌细胞增殖、克隆、迁移和细胞周期的影响  

Effect of androgen receptor-regulated long non-coding RNA ARLNC1 on proliferation,clone,migration and cell cycle of prostate cancer cells

作　　者：郑江婷 尹冶 寸淑娥 王玉明[1] ZHENG Jiangting;YIN Ye;CUN Shu'e;WANG Yuming(Department of Clinical Laboratory,the Second Affiliated Hospital of Kunming Medical University,Yunnan Kunming 650000,China)

机构地区：[1]昆明医科大学第二附属医院检验科,云南昆明650000

出　　处：《现代肿瘤医学》2024年第2期220-226,共7页Journal of Modern Oncology

基　　金：云南省科技人才和平台计划(编号:2019IC034);昆明医科大学第二附属医院院内科技计划资助项目(编号:2020yk004)。

摘　　要：目的:探讨雄激素受体(androgen receptor,AR)调控的长链非编码RNA(long non-coding RNA,lncRNA)ARLNC1对前列腺癌细胞增殖、克隆、迁移和细胞周期的影响。方法:选取正常前列腺上皮细胞株WPMY-1和前列腺癌细胞株PC3、VCaP、22RV1、DU145和LNCaP为研究对象,qRT-PCR分析lncRNA ARLNC1的表达水平;双氢睾酮(DHT)以时间和浓度依赖的方式刺激LNCaP细胞,分析lncRNA ARLNC1的转录水平;采用数据库预测和双荧光素酶报告基因系统分析雄激素受体与lncRNA ARLNC1的关系;将LNCaP细胞分为sh-NC和sh-lncRNA ARLNC1组,采用CCK-8分析细胞增殖,检测细胞克隆和迁移能力;流式细胞仪分析细胞周期变化,Western blot分析细胞周期蛋白CyclinD1、CDK6、p27的表达水平。结果:与正常前列腺上皮细胞相比,lncRNA ARLNC1在雄激素依赖性前列腺癌细胞株(LNCaP、VCaP、22RV1)中的表达水平明显增加,而在雄激素非依赖性前列腺癌细胞株(PC3、DU145)中几乎不表达(P<0.05),其中LNCaP细胞变化最显著,所以下续选用LNCaP细胞作为实验株。100 nmol/L DHT刺激LNCaP细胞0、6、12、18、24 h后,lncRNA ARLNC1的转录水平相对于对照组以时间依赖的方式逐渐升高(P<0.05);不同浓度(0、0.1、1、10、100 nmol/L)DHT刺激LNCaP细胞24 h后,lncRNA ARLNC1转录水平随浓度不断增加(P<0.05);JASPAR数据库预测到lncRNA ARLNC1启动子区域有AR结合的位点,双荧光素酶报告基因系统表明AR能结合在lncRNA ARLNC1启动子区域,并激活其转录(P<0.05);与sh-NC组相比,sh-lncRNA ARLNC1组细胞增殖、迁移和克隆能力减弱,细胞周期阻滞在S、G 2期(P<0.05);sh-NC组相对于sh-lncRNA ARLNC1组,CyclinD1、CDK6蛋白水平增加,而p27蛋白水平降低(P<0.05)。结论:AR调控的lncRNA ARLNC1作为癌基因在雄激素受体阳性的前列腺癌细胞系中高表达,能促进前列腺癌细胞的增殖、克隆、迁移和细胞周期进展。Objective:To investigate the effect of androgen receptor(AR)-regulated long non-coding RNA(lncRNA)ARLNC1 on the proliferation,clone,migration and cell cycle of prostate cancer cells.Methods:Normal prostate epithelial cell line WPMY-1 and prostate cancer cell lines PC3,VCaP,22RV1,DU145 and LNCaP were selected as the study objects.The expression level of lncRNA ARLNC1 was analyzed by qRT-PCR.Dihydrotestosterone(DHT)stimulated LNCaP cells in a time-and concentration-dependent manner to analyze the transcription level of lncRNA ARLNC1.The relationship between the androgen receptor and lncRNA ARLNC1 was analyzed by using database prediction and dual luciferase reporter system.LNCaP cells were divided into sh-NC group and sh-lncRNA ARLNC1 group.CCK-8 was used to analyze cell proliferation.The cell cloning and migration ability were detected.The cell cycle changes were analyzed by flow cytometry,and the expression levels of CyclinD1,CDK6 and p27 were analyzed by Western blot.Results:Compared with normal prostate epithelial cells,lncRNA ARLNC1 expression was significantly increased in androgen-dependent prostate cancer cell lines(LNCaP,VCaP,22RV1),but hardly expressed in androgen-independent prostate cancer cell lines(PC3,DU145)(P<0.05).LNCaP cells had the most significant changes,so LNCaP cells were selected as the experimental strain next time.After 100 nmol/L DHT stimulation of LNCaP cells for 0,6,12,18 and 24 h,the transcription level of lncRNA ARLNC1 was increased in a time-dependent manner compared with the control group(P<0.05).lncRNA ARLNC1 transcription levels increased with different concentrations(0,0.1,1,10,100 nmol/L)of DHT stimulated LNCaP cells for 24 h(P<0.05).The JASPAR database predicted that there were AR binding sites in lncRNA ARLNC1 promoter region,and the dual luciferase reporter system showed that AR could bind to lncRNA ARLNC1 promoter region and activate its transcription(P<0.05).Compared with sh-NC group,the cell proliferation,migration and cloning ability of sh-lncRNA ARLNC1 group were weakene

关 键 词：前列腺癌 长链非编码RNA 雄激素受体 增殖 克隆 迁移 细胞周期 

分 类 号：R737.25[医药卫生—肿瘤]