RNA干扰沉默B细胞特异的莫洛尼白血病病毒插入位点1基因对前列腺癌PC-3细胞增殖和凋亡的影响  

Effects of silencing B-cell specific moloney leukemia virus insertion site 1 gene by RNA inerferenceon proliferation and apoptosis of prostate cancer cell line PC-3

作　　者：詹以安[1] 李煜[1] 傅龙龙[1] 刘伟鹏[1] 傅斌[1] 王共先[1] 

机构地区：[1]南昌大学第一附属医院泌尿外科研究所,南昌330006

出　　处：《中华实验外科杂志》2013年第3期584-586,共3页Chinese Journal of Experimental Surgery

基　　金：江西省研究生创新专项资金资助项目（YC2011-B009）

摘　　要：目的观察RNA干扰沉默B细胞特异性莫洛尼白血病病毒插入位点1（Bmi—1）基因对前列腺癌PC-3细胞增殖和凋亡的影响。方法PC-3细胞随机分为3组，分别转染Bmi-1靶序列干扰RNA（Bmi．1-siRNA）、无义对照siRNA、空白脂质体。48h后收集细胞，逆转录-聚合酶链反应（RT．PCR）、Westernblot检测Bmi-1mRNA和蛋白表达水平。96孔板噻唑蓝（MTT）法检测细胞增殖活力，流式细胞仪检测凋亡率。结果PC．3细胞经Bmi—l—siRNA干扰后Bmi-1mRNA和蛋白质表达水平下调（P〈0．05）。与对照组比较，实验组细胞增殖明显受到抑制（P〈0．05）。实验组细胞凋亡率为（23．77±2．23）％，与空白对照组[（7．05±0．88）％]和阴性对照组[（6．89±I．02）％]比较明显增高（P〈0．05）。结论Bmi．1．siRNA能通过沉默靶基因Bmi—l表达抑制前列腺癌PC-3细胞增殖并促进细胞凋亡。Objective To investigate the effect of silencing B-cell specific moloney leukemia virus insertion site 1 （ BMI-1 ） gene by RNA interference on the proliferation and apoptosis of prostate cancer cell line PC-3. Methods PC-3 cells were divided randomly into 3 groups ： experimental group, negative con- trol group and blank control group, which was transfected with Bmi-l-siRNA, nonsence-siRNA and lipo- fectamine respectively by LipofectamineTM 2000. The cells were collected at 48 h after transfection. The ex- pression of Bmi-I mRNA and protein in the transfected cells was detected by using reverse transcription- polymerase chain reaction （RT-PCR） and Western blotting. Cell growth was measured by using methyl thi- azol tetrazolium （MTr） assay, and cell apoptosis rate was examined by using flow cytometry. Results The expression of Bmi-1 mRNA and protein in PC-3 cells transfected with Bmi-l-siRNA was markedly down-regu- lated （P 〈 0. 05 ）, As compared with the blank and negative control groups, the proliferation of PC-3 cells transfected with Bmi-l-siRNA was significantly decreased （P 〈 0.05 ）. The apoptosis rate was （23.77±2. 23）% in PC-3 cells transfected with Bmi-l-siRNA, significantly higher than that in blank control group [ （7.03 ±0.88） % ] and negative control group [ （6. 89 ±1.02） % ] （P 〈 0.05 ）. Conclusion Down-regu- lation of Bmi-1 gene can inhibit the proliferation and promote apoptosis of PC-3 ceils, and Bmi-1 gene may serve as an effective tarzet for the gene therapy of prostate cancer.

关 键 词：前列腺肿瘤 B细胞特异性莫洛尼白血病病毒插入位点1基因 RNA干扰 细胞增殖 

分 类 号：R737[医药卫生—肿瘤]