稳定沉默Raptor表达的前列腺癌DU145细胞株的构建  

Construction of the prostate cancer cell line DU145 with Raptor-shRNA

作　　者：潘腾飞[1] 陈先国[1] 梁朝朝[1] 

机构地区：[1]安徽医科大学第一附属医院泌尿外科,合肥230022

出　　处：《安徽医科大学学报》2013年第4期346-348,共3页Acta Universitatis Medicinalis Anhui

基　　金：教育部博士点基金(编号:20113420110003);安徽省自然科学基金(编号:1208085MH138)

摘　　要：目的应用RNA干扰技术降低前列腺癌DU145细胞株Raptor基因表达水平,并建立稳定转染细胞株。方法针对Raptor基因设计合成siRNA,鉴定、测序正确后转染293T细胞观察基因表达情况;构建Raptor-shRNA慢病毒载体;进行Raptor-shRNA慢病毒包装及滴度测定;筛选稳定表达Raptor-shRNA的前列腺癌DU145细胞株;Western blot检测稳定转染细胞中的Raptor表达水平。结果 Raptor-shR-NA成功转染前列腺癌DU145细胞后,通过嘌呤霉素筛选,获得了Raptor基因沉默的DU145细胞株,Western blot检测证实该细胞株的Raptor表达水平明显低于对照组(P<0.01)。结论成功构建稳定沉默Raptor表达的前列腺癌DU145细胞株,为后续研究奠定了基础。Objective To establish a prostate cancer cell line DU145 with raptor stably silenced by RNAi for for- ther investigation of raptor in prostate cancer. Methods The sequence of siRNA targeting to raptor was designed and synthesized. Followed by transfection of 293T cells to observe the expression of raptor, a lentiviral vector was constructed. Packaging of lentiviral vector carrying the raptor-shRNA was performed and the expression was evalua- ted after screening. Results A lentiviral vector carrying raptor-shRNA was successfully constructed and transfeeted into prostate cancer cell line DU145. The DU145 cell line was obtained with raptor silencing by puromycin screen- ing. The expression of raptor in prostate cancer cell line DU145 was remarkably decreased （P 〈 0. 01 ）. Conclu- sion The prostate cancer cell line DU145 with raptor has been successfully silenced by RNAi, which may lay the foundation for further approach in prostate cancer.

关 键 词：SHRNA RAPTOR 前列腺癌 DU145 

分 类 号：R737.25[医药卫生—肿瘤]