Guan Han, Ling Zhi-Xin, Fang Fang, Mao Li-Kai, You Zong-Hao, Wang Can, Chen Shu-Qiu, Xu Bin, Chen Ming
Department of Urology, The First Hospital of Bengbu Medical College,Bengbu Medical College, Bengbu, Anhui 233000, China.
Graduate School of Medical College,Southeast University, Nanjing, Jiangsu 210009, China..
Zhonghua Nan Ke Xue. 2018 Feb;24(2):116-121.
To establish enzalutamide-resistant human prostate cancer cell lines and screen out the lncRNA and mRNA expression profiles associated with enzalutamide resistance.
Human prostate cancer cell lines LNCAP and C4-2B were cultured with 10 μmol/L enzalutamide for 6 months in vitro for the establishment of enzalutamide-resistant subclones LNCAP-ENZA and C4-2B-ENZA. The IC50 value and enzalutamide resistance index of each cell line were examined by MTT assay, the expressions of enzalutamide-related genes FL-AR, AR-V7 and HnRNPA1 were determined by Western blot, and the lncRNA and mRNA differential expressions of C4-2B and C4-2B-ENZA were detected by high-throughout lncRNA microarray.
Compared with LNCAP and C4-2B, the IC50 values of enzalutamide-resistant subclones LNCAP-ENZA (60.83 μmol/L) and C4-2B-ENZA (88.32 μmol/L) were increased significantly (P < 0.05) and the enzalutamide-resistance indexes of the LNCAP-ENZA and C4-2B-ENZA cells were 4.94 and 4.67, respectively. The expressions of AR-V7 and HnRNPA1 were markedly up-regulated in the LNCAP-ENZA and C4-2B-ENZA cells as compared with those in the LNCAP and C4-2B cells, but that of FL-AR showed no significant change. A total of 1 440 lncRNAs and 1 236 mRNAs were identified as differentially expressed in the C4-2B-ENZA cells.
Enzalutamide -resistant human prostate cancer cell subclones LNCAP-ENZA and C4-2B-ENZA were successfully established and enzalutamide resistance-associated lncRNA and mRNA were identified, which may provide some molecular evidence for the management of enzalutamide-resistant human prostate cancer.
建立恩杂鲁胺耐药的人前列腺癌细胞系，并筛选出与恩杂鲁胺耐药相关的lncRNA和mRNA表达谱。
将人前列腺癌细胞系LNCAP和C4-2B用10 μmol/L恩杂鲁胺体外培养6个月，以建立恩杂鲁胺耐药亚克隆LNCAP-ENZA和C4-2B-ENZA。通过MTT法检测各细胞系的IC50值和恩杂鲁胺耐药指数，采用蛋白质免疫印迹法检测恩杂鲁胺相关基因FL-AR、AR-V7和HnRNPA1的表达，通过高通量lncRNA芯片检测C4-2B和C4-2B-ENZA的lncRNA和mRNA差异表达。
与LNCAP和C4-2B相比，恩杂鲁胺耐药亚克隆LNCAP-ENZA（60.83 μmol/L）和C4-2B-ENZA（88.32 μmol/L）的IC50值显著升高（P < 0.05），LNCAP-ENZA和C4-2B-ENZA细胞的恩杂鲁胺耐药指数分别为4.94和4.67。与LNCAP和C4-2B细胞相比，LNCAP-ENZA和C4-2B-ENZA细胞中AR-V7和HnRNPA1的表达明显上调，但FL-AR的表达无显著变化。共鉴定出1440个lncRNA和1236个mRNA在C4-2B-ENZA细胞中差异表达。
成功建立了恩杂鲁胺耐药的人前列腺癌细胞亚克隆LNCAP-ENZA和C4-2B-ENZA，并鉴定出与恩杂鲁胺耐药相关的lncRNA和mRNA，这可能为恩杂鲁胺耐药的人前列腺癌的治疗提供一些分子证据。
