本文選題：前列腺癌　切入點(diǎn)：HuR　出處：《中華腫瘤防治雜志》2016年05期

【摘要】：目的前列腺癌在我國(guó)的發(fā)病率及病死率逐年上升,但其發(fā)病機(jī)制尚未明確,本研究從轉(zhuǎn)錄后水平初步探討RNA結(jié)合蛋白HuR參與調(diào)控前列腺癌細(xì)胞活力的機(jī)制。方法免疫熒光檢測(cè)前列腺增生細(xì)胞(BPH)及前列腺癌細(xì)胞(PC3)中HuR蛋白的定位,蛋白質(zhì)印跡法檢測(cè)HuR及環(huán)氧合酶-2(cyclooxygenase,COX-2)蛋白在細(xì)胞內(nèi)的表達(dá),MTT法檢測(cè)細(xì)胞活力,脂質(zhì)體轉(zhuǎn)染法轉(zhuǎn)染質(zhì)粒及干擾片段,qPCR檢測(cè)干擾效率,RNA-pull down實(shí)驗(yàn)驗(yàn)證HuR對(duì)COX-2的調(diào)控。結(jié)果 HuR在BPH細(xì)胞主要定位于細(xì)胞核,在PC3細(xì)胞中主要定位于胞質(zhì)。PC3細(xì)胞中HuR表達(dá)水平為1.699±0.011,高于BPH細(xì)胞的0.654±0.028,差異有統(tǒng)計(jì)學(xué)意義,t=58.67,P0.001。PC3細(xì)胞中過(guò)表達(dá)HuR后,轉(zhuǎn)染空質(zhì)粒組細(xì)胞活力為1.038±0.117,轉(zhuǎn)染HuR表達(dá)質(zhì)粒后細(xì)胞活力為1.838±0.057,差異有統(tǒng)計(jì)學(xué)意義,t=10.656,P0.001。干擾HuR表達(dá)后,轉(zhuǎn)染siNC組細(xì)胞活力為1.254±0.095,轉(zhuǎn)染siHuR干擾片段組細(xì)胞活力為0.66±0.102,t=7.383,P=0.002;BPH組和PC3組COX-2表達(dá)水平分別為0.449±0.055和1.066±0.068,t=12.147,P0.001;過(guò)表達(dá)COX-2后PC3細(xì)胞活力明顯增加,轉(zhuǎn)染空質(zhì)粒組細(xì)胞活力為1.296±0.114,轉(zhuǎn)染COX-2表達(dá)質(zhì)粒后細(xì)胞活力為1.954±0.062,t=18.062,P0.001。干擾COX-2表達(dá)后,PC3細(xì)胞活力明顯降低,轉(zhuǎn)染siNC組細(xì)胞活力為1.233±0.145,轉(zhuǎn)染siCOX-2干擾片段后細(xì)胞活力為0.661±0.096,t=5.688,P=0.005。HuR可以結(jié)合于COX-2的3′-UTR并上調(diào)COX-2蛋白表達(dá),轉(zhuǎn)染空質(zhì)粒組COX-2蛋白表達(dá)水平為0.638±0.067,轉(zhuǎn)染HuR表達(dá)質(zhì)粒組為1.703±0.022,t=26.26,P0.001。結(jié)論HuR參與調(diào)控前列腺癌細(xì)胞活力可能是通過(guò)上調(diào)COX-2表達(dá)而實(shí)現(xiàn)。
[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 鹵0.011), which was higher than that in BPH cells (0.654 鹵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 鹵0.117, and that of HuR expression plasmid was 1.838 鹵0.057. The difference was statistically significant (P 0.001).騫叉壈HuR琛ㄨ揪鍚，

本文編號(hào)：1703716