RNA结合蛋白HuR调控前列腺癌细胞活力机制的研究

发布时间：2018-04-03 05:03

  本文选题：前列腺癌　切入点：HuR　出处：《中华肿瘤防治杂志》2016年05期

【摘要】：目的前列腺癌在我国的发病率及病死率逐年上升,但其发病机制尚未明确,本研究从转录后水平初步探讨RNA结合蛋白HuR参与调控前列腺癌细胞活力的机制。方法免疫荧光检测前列腺增生细胞(BPH)及前列腺癌细胞(PC3)中HuR蛋白的定位,蛋白质印迹法检测HuR及环氧合酶-2(cyclooxygenase,COX-2)蛋白在细胞内的表达,MTT法检测细胞活力,脂质体转染法转染质粒及干扰片段,qPCR检测干扰效率,RNA-pull down实验验证HuR对COX-2的调控。结果 HuR在BPH细胞主要定位于细胞核,在PC3细胞中主要定位于胞质。PC3细胞中HuR表达水平为1.699±0.011,高于BPH细胞的0.654±0.028,差异有统计学意义,t=58.67,P0.001。PC3细胞中过表达HuR后,转染空质粒组细胞活力为1.038±0.117,转染HuR表达质粒后细胞活力为1.838±0.057,差异有统计学意义,t=10.656,P0.001。干扰HuR表达后,转染siNC组细胞活力为1.254±0.095,转染siHuR干扰片段组细胞活力为0.66±0.102,t=7.383,P=0.002;BPH组和PC3组COX-2表达水平分别为0.449±0.055和1.066±0.068,t=12.147,P0.001;过表达COX-2后PC3细胞活力明显增加,转染空质粒组细胞活力为1.296±0.114,转染COX-2表达质粒后细胞活力为1.954±0.062,t=18.062,P0.001。干扰COX-2表达后,PC3细胞活力明显降低,转染siNC组细胞活力为1.233±0.145,转染siCOX-2干扰片段后细胞活力为0.661±0.096,t=5.688,P=0.005。HuR可以结合于COX-2的3′-UTR并上调COX-2蛋白表达,转染空质粒组COX-2蛋白表达水平为0.638±0.067,转染HuR表达质粒组为1.703±0.022,t=26.26,P0.001。结论HuR参与调控前列腺癌细胞活力可能是通过上调COX-2表达而实现。
[Abstract]:Objective the incidence and mortality of prostate cancer in China have been increasing year by year, but the pathogenesis of prostate cancer has not been clarified. In this study, the mechanism of RNA binding protein HuR involved in the regulation of prostate cancer cell viability was preliminarily explored at the post-transcriptional level.Methods Immunofluorescence assay was used to detect the localization of HuR protein in prostatic hyperplasia cells (BPH) and prostate cancer cell line (PC3). Western blotting was used to detect the expression of HuR and cyclooxygenase cyclooxygenase (COX-2) protein in the cells.Detection of interference efficiency by Liposome transfection of plasmid and interference fragment down the regulation of HuR on COX-2 was verified by RNA-pull down experiment.Results the expression of HuR was mainly located in the nucleus of BPH cells and in the cytoplasm of PC3 cells (1.699 卤0.011), which was higher than that in BPH cells (0.654 卤0.028). The difference was statistically significant after the expression of HuR in P0.001.PC3 cells.The cell viability of empty plasmid group was 1.038 卤0.117, and that of HuR expression plasmid was 1.838 卤0.057. The difference was statistically significant (P 0.001).骞叉壈HuR琛ㄨ揪鍚，

本文编号：1703716