靶向人乳腺癌HPSE基因的shRNA慢病毒表达载体的构建及鉴定  

Construction and identification of lentivirus-based vectors of short hairpin RNA targeting human HPSE in breast cancer

作　　者：陈国利[1,2] 王烈[1,2] 

机构地区：[1]福建医科大学福总临床医学院,福建福州350025 [2]南京军区福州总医院普外科,福建福州350025

出　　处：《中国现代普通外科进展》2011年第1期24-28,共5页Chinese Journal of Current Advances in General Surgery

基　　金：南京军区"十一五"科研规划重点课题(07Z038)

摘　　要：目的:应用RNA干扰技术构建HPSE shRNA重组慢病毒载体,并筛选出最显著抑制HPSE表达的shRNA干扰序列。方法:体外化学合成针对HPSE基因的siRNA序列,在慢病毒载体介导下转染MDA-MB-231细胞,实时定量PCR检测HPSE mRNA表达水平。结果:HPSE shRNA转染MDA-MB-231细胞72 h后,RT-PCR结果表明:所设计的siRNA1组、siRNA4组针对不同靶点的shRNA与对照组相比均可有效抑制HPSE的表达,且成功筛选出最有效抑制HPSE表达的shRNA4序列(P<0.05)。结论:成功构建了HPSE shRNA重组慢病毒载体,其转染乳腺癌MDA-MB-231细胞后显著抑制了HPSE的表达,为进一步探讨乳腺癌的侵袭转移机制及肿瘤的基因治疗提供了实验基础。Objective： To construct five short hairpin RNA（shRNA） interference recombinant lentivirus-based vectors of human HPSE gene by using RNA interference,and detect the strongest RNAi effect of HPSE shRNA sequence.Methods：Chemically synthesized small interfering RNA（siRNA） sequences targeting HPSE gene in vitro was transfected into human breast cancer MDA-MB-231 cells by lentivirus-based vectors.The expression levels of HPSE mRNA were detected by real time PCR（RT-PCR）.Results：RT-PCR results showed that the expressions levels of HPSE mRNA of MDA-MB-231 cells were effectively decreased by siRNA1 and siRNA4 than the control group,and successfully elected the effectively decreasest shRNA4 sequence targeting HPSE gene at 72 hours after transfection（P0.05）.Conclusion： The results indicate that the HPSE shRNA recombinant lentivirus vectors was successfully constructed,and the shRNA can significantly inhibit the expression level of HPSE.This finding could provide an experimental basis for further study on invasiveness,metastasis of breast cancer,and its application in tumour gene therapy.

关 键 词：乳腺肿瘤 HPSE SHRNA RNA干扰 慢病毒载体 

分 类 号：R737.9[医药卫生—肿瘤]