miR-539-5p对雄激素非依赖性前列腺癌细胞阿帕鲁胺敏感性及恶性表型的影响及机制研究  

Effect of miR-539-5p on ARN-509 sensitivity and malignant phenotype of androgen independent prostate cancer cells and its mechanism

作　　者：杨占锋[1] 王雷[1] 邱晓东[1] 李建华[1] 朱照伟[2] Yang Zhanfeng;Wang Lei;Qiu Xiaodong;Li Jianhua;Zhu Zhaowei(Department of Urology,Shangqiu First people’s Hospital,Shangqiu 476100,China;Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]商丘市第一人民医院泌尿外科,商丘476100 [2]郑州大学第一附属医院泌尿外科,郑州450052

出　　处：《中华内分泌外科杂志》2022年第3期320-324,共5页Chinese Journal of Endocrine Surgery

基　　金：国家自然科学基金(81702503)。

摘　　要：目的研究miR-539-5p对雄激素非依赖性前列腺癌细胞C4-2B阿帕鲁胺(ARN-509)敏感性及恶性表型的影响及相关机制。方法获取去势抵抗性前列腺癌、去势敏感性前列腺癌及良性前列腺组织,并选取处于对数生长期C4-2B前列腺癌细胞,传代扩增后,采用miR-539-5p质粒对C4-2B细胞系进行转染,共分为空白组,转染组(miR-539-5p质粒)及对照组(对照质粒)。qPCR检测组织及3组细胞中miR-539-5p、AR及热休克因子结合蛋白1(heat shock factor binding protein 1,HSBP1)基因的表达含量;Western blot检测3组细胞雄激素受体AR及HSBP1的蛋白表达含量;Transwell实验检测3组细胞的侵袭迁移能力;CCK-8法检测3组细胞的增殖能力及雄激素受体拮抗剂ARN-509的半抑制浓度(IC_(50));平板克隆实验检测3组细胞的克隆形成能力。结果组织qPCR提示:良性前列腺组织、前列腺癌去势敏感患者肿瘤组织及前列腺癌去势抵抗患者肿瘤组织miR-539-5p的表达分别为0.29±0.04、0.17±0.02、0.07±0.01,AR的表达分别为0.13±0.02、0.28±0.04、0.79±0.11,HSBP1的表达分别为0.20±0.03、0.38±0.04、0.72±0.11。相比良性前列腺组织和前列腺癌组织,前列腺癌去势抵抗患者的组织中AR及HSBP1基因的表达含量较高,miR-539-5p表达较低(P<0.001)。细胞qPCR提示:空白组、对照组及转染组miR-539-5p表达分别为1.00±0.09、1.07±0.11、7.19±0.51,AR的表达分别为1.00±0.10、1.03±0.14、0.51±0.08,HSBP1的表达分别为1.00±0.10、0.96±0.12、0.97±0.11。转染组细胞的miR-539-5p表达显著高于对照组及空白组,AR基因表达含量明显低于对照组及空白组,HSBP1基因表达水平无显著差异。Western blot显示:空白组、对照组及转染组AR的蛋白表达分别为1.00±0.10、1.12±0.22、0.72±0.16,HSBP1的蛋白表达分别为1.00±0.10、0.94±0.18、0.48±0.11,转染组细胞的AR及HSBP1的蛋白表达明显少于对照组及空白组。Transwell实验显示:转染组侵袭及迁移细�Objective To study the effect of miR-539-5p on apalutamide(ARN-509)sensitivity and malignant phenotype of androgen independent prostate cancer cell line C4-2B and related mechanisms.Methods Castrated resistant prostate cancer,castrated sensitive prostate cancer and benign prostate tissue were obtained.C4-2B cell lines were divided into blank group,transfection group(miR-539-5p plasmid)and control group(control plasmid).qPCR was used to detect the expression of miR-539-5p,androgen receptor(AR)and HSBP1 in the tissues and 3 group of cells.The protein expressions of AR and HSBP1 were detected by western blot.Transwell assay was used to detect the invasion and migration ability of three groups of cells.CCK-8 assay was used to detect the proliferation ability and semi-inhibitory concentration(IC_(50))of AR antagonist ARN-509.The colony forming ability of the three groups of cells was detected by plate cloning experiment.Results Tissue-qPCR indicated that,in the benign prostate tissue,tumor tissue of castration sensitive patients and tumor tissue of castration resistant patients,the expressions of miR-539-5p were 0.29±0.04,0.17±0.02 and 0.07±0.01,the expressions of AR were 0.13±0.02,0.28±0.04 and 0.79±0.11,and the expressions of HSBP1 were 0.20±0.03,0.38±0.04 and 0.72±0.11,respectively.Compared with benign prostate tissue and prostate cancer tissue,the expression of AR and HSBP1 gene was higher in prostate cancer tissues with castration resistance,and the expression of miR-539-5p was lower.Cell-qPCR demonstrated that the expressions of miR-539-5p in blank group,control group and transfection group were 1.00±0.09,1.07±0.11 and 7.19±0.51,the expressions of AR were 1.00±0.10,1.03±0.14 and 0.51±0.08,and the expressions of HSBP1 were 1.00±0.10,0.96±0.12 and 0.97±0.11.The expression of miR-539-5p in the transfection cells was significantly higher than that in the control group and the blank group,the expression of AR gene was significantly lower than that in the control group and the blank group,and there w

关 键 词：miR-539-5p 热休克因子结合蛋白1 前列腺癌 去势抵抗 

分 类 号：R697+.3[医药卫生—泌尿科学]