癌相关成纤维细胞外泌体微小RNA-520e通过Hippo通路增强前列腺癌PC-3细胞多西他赛耐药性  

Cancer-associated fibroblast-secreted exosomal microRNA-520e induces docetaxel resistance in prostate cancer PC-3 cell through the Hippo signaling pathway

作　　者：陈航 黄琦 陈佳茵 林婷婷[1] 林云知 孙雄林[1] 郑清水[1] 魏勇[1] 许宁[1] 薛学义[1] Chen Hang;Huang Qi;Chen Jiayin;Lin Tingting;Lin Yunzhi;Sun Xionglin;Zheng Qingshui;Wei Yong;Xu Ning;Xue Xueyi(Departments of Urology,the First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China)

机构地区：[1]福建医科大学附属第一医院泌尿外科,福州350005

出　　处：《中华实验外科杂志》2022年第2期308-311,共4页Chinese Journal of Experimental Surgery

基　　金：福建省自然科学基金(2019J01445)。

摘　　要：目的探讨癌相关成纤维细胞(CAFs)来源外泌体微小RNA(miR)-520e对前列腺癌细胞多西他赛耐药性的影响及其机制。方法 2019年5月至2019年12月, 从福建医科大学附属第一医院泌尿外科前列腺癌手术患者癌旁组织获取原代CAFs, 提取并鉴定CAFs外泌体后, 将其与PC-3细胞共培养, 细胞计数试剂盒(CCK-8)法检测其在高浓度(30 nmol/L)、低浓度(5 nmol/L)多西他赛条件下细胞增殖能力, 计算细胞抑制率。构建高表达及低表达miR-520e的CAFs, 提取外泌体后与PC-3细胞共培养, 检测其对细胞增殖能力的影响, 并计算细胞抑制率。进一步, 通过蛋白质印迹法(Western blot)检测Hippo信号通路标志蛋白水平, 验证此通路是否参与外泌体miR-520e调控PC-3细胞多西他赛耐药过程。使用方差分析和独立样本t检验比较组间差异。结果 PC-3细胞抑制率与多西他赛浓度呈正比(Pearsonr=0.826, P<0.05);加入CAFs来源外泌体后, 高、低浓度多西他赛组细胞抑制率分别为(50.510±2.389)%和(28.233±5.441)%, 均低于对照组(t=12.010, P<0.05和t=5.614, P<0.05)。高、低表达miR-520e外泌体处理组中, PC-3细胞抑制率分别为(26.355±1.539)%和(72.965±1.777)%, 与对照组[(59.856±2.570)%]比较, 差异均有统计学意义(t=19.370, P<0.05与t=7.270, P<0.05)。此外, 高表达miR-520e外泌体处理组LATS1表达水平低于对照组(0.481±0.005比1.765±0.015, t=139.200, P<0.05), YAP表达高于对照组(0.932±0.001比0.688±0.005, t=81.850, P<0.05);而低表达miR-520e外泌体处理组则相反, LATS1表达高于对照组(2.370±0.050比1.765±0.015, t=16.770, P<0.05), YAP表达则低于对照组(0.371±0.002比0.688±0.005, t=97.190, P<0.05)。结论 CAFs来源外泌体miR-520e通过调控Hippo通路增强前列腺癌PC-3细胞对多西他赛的耐药。Objective To investigate the effects and mechanism of cancer-associated fibroblasts(CAFs)-derived exosomal microRNA(miR)-520e on resistance of prostate cancer cells towards docetaxel.Methods CAFs were extracted from paracancerous tissue samples derived from patients who underwent radical prostatectomy in Department of Urology,the First Affiliated Hospital of Fujian Medical,between May 2019 and December 2019.Exosomes derived from CAFs were extracted and identified followed by co-culture with PC-3 cells.The proliferation ability of PC-3 in high(30 nmol/L)or low(5 nmol/L)concentration of docetaxel was detected using cell counting kit-8(CCK-8)assay,and the growth inhibition rates were calculated.CAFs with high or low miR-520e expression level were constructed and co-cultured with the PC-3 cells,and then the proliferation ability and the growth inhibition rate of PC-3 cells were assessed.Western blotting was performed to detect the expression level of Hippo signaling pathway markers,and to explore whether exosomal miR-520e regulated resistance of PC-3 cells towards docetaxel via this signaling pathway.ANOVA and Student t test were used for statistical analysis.Results We observed a positive correlation between the growth inhibition rate and the drug concentration of docetaxel(Pearson r=0.826,P<0.05).After adding CAFs-derived exosomes,the inhibition rate of PC-3 cells in high and low concentration of docetaxel groups was(50.510±2.389)%and(28.233±5.441)%respectively,significantly lower than that without adding exosomes(t=12.010,P<0.05 and t=5.614,P<0.05).The inhibition rates of PC-3 cells treated with high and low expression miR-520e exosomes were(26.355±1.539)%and(72.965±1.777)%,respectively.The differences were statistically significant(t=19.370,P<0.05 and t=7.270,P<0.05),when compared with the control group[(59.856±2.570)%].Compared with the control group,the expression level of LATS1 in the group treated with exosomes overexpressing miR-520e was lower(0.481±0.005 vs.1.765±0.015,t=139.200,P<0.05),and that of Y

关 键 词：前列腺癌 外泌体 化疗耐药 癌相关成纤维细胞 

分 类 号：R737.25[医药卫生—肿瘤]