交感神经β2肾上腺素受体促进前列腺癌转移并抵抗肿瘤细胞的凋亡  

Sympathetic β2 adrenergic receptor promotes prostate cancer metastasis and resists tumor cell apoptosis

作　　者：张咪 王千慧 陈津滢 尹青青 徐晨[1] ZHANG Mi;WANG Qianhui;CHEN Jinying;YIN Qingqing;XU Chen(Institution of Life Science,Chongqing Medical University,Chongqing,400016,China)

机构地区：[1]重庆医科大学生命科学研究院

出　　处：《第三军医大学学报》2019年第21期2051-2060,共10页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2009BA5081)~~

摘　　要：目的探讨交感神经β2肾上腺素受体(β2 adrenergic receptor,β2AR)对前列腺癌细胞迁移、侵袭以及凋亡的影响。方法通过UALCAN数据库检索分析β2AR在人前列腺癌组织及癌旁组织中的基因表达水平及与患者年龄的相关性;通过免疫组织化学染色分析β2AR在29例前列腺癌组织中的蛋白表达水平。体外培养人前列腺癌细胞系PC-3、LNCaP,β2AR激动剂异丙肾上腺素(ISO,2μmol/L,24 h或1、12、24 h),β2AR抑制剂普萘洛尔(PRO,1μmol/L,24 h)处理细胞;β2AR RNAi慢病毒转染PC-3、LNCaP细胞,Western blot及RT-PCR分别检测β2AR的蛋白及mRNA水平。Transwell实验检测细胞迁移及侵袭。Western blot检测上皮间充质转化(epithelial mesenchymal transformation,EMT)相关标记物E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和凋亡相关蛋白Caspase3、cleaved-Caspase3及PARP的表达;流式细胞仪检测凋亡率;透射电镜观测细胞凋亡情况。结果 UALCAN数据库检索结果显示β2AR在前列腺癌患者组织中的基因表达高于正常前列腺组织(P<0.05);β2AR在41~60岁与61~80岁患者组间的表达差异不显著。29例前列腺癌患者组织的免疫组化结果显示,高风险患者(Gleason评分≥7)组织中β2AR蛋白水平高于低风险患者(Gleason评分<7)(P<0.001);细胞迁移及侵袭实验中,ISO处理组细胞迁移和侵袭增加(P<0.01),E-cadherin表达下调,N-cadherin和Vimentin表达上调(P<0.05);而PRO处理组及下调β2AR的表达后,细胞迁移和侵袭减少(P<0.01),E-cadherin表达上调,N-cadherin和Vimentin表达下调(P<0.05)。细胞凋亡实验中,ISO处理组细胞凋亡率降低,Caspase3、cleaved-Caspase3、PARP的表达明显降低(P<0.05),而PRO处理组及下调β2AR的表达细胞凋亡率明显增高,且Caspase3、cleaved-Caspase3、PARP的表达增高(P<0.05)。结论β2AR能够促进细胞EMT的形成,增加前列腺癌细胞的迁移、侵袭能力,并且通过下调凋亡相关蛋白的表达,对细胞Objective To investigate the effects of sympathetic β2 adrenergic receptor(β2 AR) on the migration, invasion and apoptosis of prostate cancer cells. Methods The gene expression levels of β2 AR in human prostate cancer tissues and adjacent tissues were analyzed by UALCAN database retrieval. Then, the protein level of β2 AR in human prostate cancer tissues from 29 cases was analyzed by immunohistochemical assay. β2 AR agonist isoproterenol(ISO, 2 μmol/L, 24 h, or 1, 12 and 24 h) and β2 AR inhibitor propranolol(PRO, 1 μmol/L, 24 h) were used to treat human prostate cancer cell lines PC-3 and LNCaP, respectively, and lentiviral vectors containing β2 AR RNAi were transfected to the cell lines. The protein and mRNA levels of β2 AR were detected by Western blotting and RT-PCR, respectively. Cell migration and invasion were detected by Transwell assay. Western blot assay was performed to detect the expression of epithelial mesenchymal transformation(EMT)-related markers, E-cadherin, N-cadherin and vimentin, and apoptosis-related proteins, Caspase3, cleaved-Caspase3 and PARP. Flow cytometry was used to detect cell apoptosis. Transmission electron microscopy was used to observe the apoptotic cells. Results The results of UALCAN database retrieval showed that the expression level of β2 AR were significantly higher in prostate cancer tissues than normal prostate tissues(P<0.05), and the expression showed no significant difference between the patients of 41~60 and 61~80 years old. Among the 29 patients, those with high risk(Gleason score≥7) showed higher expression of β2 AR than low risk(Gleason score <7) patients(P<0.001). In cells migration and invasion assays, ISO treatment promoted the migration and invasion of cells(P<0.01), down-regulated the expression of E-cadherin, and up-regulated N-cadherin and vimentin levels(P<0.05), while PRO treatment and β2 AR knockdown reduced the expression of β2 AR, inhibited the migration and invasion(P<0.01), enhanced the expression of E-cadherin, and decreased the express

关 键 词：前列腺癌 Β2肾上腺素受体 转移 凋亡 

分 类 号：R73-37[医药卫生—肿瘤] R730.23[医药卫生—临床医学]