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顺铂诱导自噬和MAPK信号调控前列腺癌细胞耐药的机制研究
作者:刘强1  木拉提· 热夏提1  拜合提亚· 阿扎提1  杨颖2  王文光1 
单位:1. 新疆医科大学第一附属医院 泌尿中心, 新疆 乌鲁木齐 830054;
2. 新疆医科大学第一附属医院 肿瘤中心, 新疆 乌鲁木齐 830054
关键词:顺铂 前列腺癌细胞 自噬 丝裂原活化蛋白激酶信号 耐药 
分类号:R737.25
出版年·卷·期(页码):2018·46·第十二期(1325-1331)
摘要:

目的:探讨自噬和丝裂原活化蛋白激酶(MAPK)信号在前列腺癌细胞顺铂(CDDP)化疗后耐药性获得过程中所发挥的功能。方法:通过细胞克隆分析实验检测不同CDDP浓度下的细胞存活情况以确定最佳处理浓度及耐药细胞株,Western blotting检测自噬和MAPK信号相关蛋白表达含量变化,逆转录-聚合酶链反应(RT-PCR)检测自噬相关基因转录水平变化,AnnexinV-FITC及Z-VAD-FMK检测细胞凋亡情况,利用药物抑制自噬和MAPK信号通路探讨两者对前列腺癌细胞耐药性获得的贡献。结果:细胞克隆分析实验结果显示,7.5 μg·ml-1 CDDP诱导显著的细胞凋亡,通过连续10 d的药物筛选获得耐药细胞株,显微镜下观察到前列腺癌细胞获得耐药性后细胞形态发生变化,同时DNA损伤修复的关键因子PARP蛋白表达上调;RT-PCR及Western blotting结果显示CDDP激活了前列腺癌细胞中的自噬过程,ATG5和LC3B-Ⅱ蛋白及mRNA表达水平均上调(P<0.05);AnnexinV-FITC及Z-VAD-FMK实验结果显示抑制了自噬后细胞凋亡显著增加(P<0.05),前列腺癌细胞恢复了对CDDP的敏感性,且此过程有MAPK信号的参与。结论:自噬和MAPK信号在前列腺癌细胞CDDP耐药过程中发挥重要作用,这一结果可对改善前列腺癌患者的治疗策略提供新思路。

Objective: To investigate the role of autophagy and mitogen-activated protein kinase (MAPK) signaling in acquired resistance of prostate cancer cell after cisplatin (CDDP) chemotherapy. Methods:Cell cloning assay was used to determine the cell survival of different CDDP concentrations, and confirm the best treatment concentration and drug-resistant cell line; Western blotting was used to detect the expression of autophagy and MAPK signaling proteins; Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the transcription level of autophagy-related genes. AnnexinV-FITC and Z-VAD-FMK were used to detect apoptosis. The role of drugs inhibit autophagy and MAPK signaling pathway on acquired resistance of prostate cancer cell was explored. Results:The results of cell clone analysis showed that 7.5 μg·ml-1 CDDP concentration induced significant apoptosis, drug-resistant cell lines were obtained by drug screening for 10 consecutive days, and the morphological changes of prostate cancer cells after drug resistance were observed under microscope. The expression of DNA repair key factor PARP protein was upregulated, RT-PCR and Western blotting results showed that CDDP activated autophagy in prostate cancer cells, and upregulated the protein and mRNA levels of ATG5 and LC3B-Ⅱ(P<0.05).The results of Annexin V-FITC and Z-VAD-FMK showed that apoptosis increased significantly when autophagy was inhibited (P<0.05), prostate cancer cells recovered their sensitivity to CDDP, and MAPK signaling was involved in this process. Conclusion:Autophagy and MAPK signaling play an important role in the process of CDDP resistance of prostate cancer cells, which may provide new ideas for improving the treatment strategies of prostate cancer patients.

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