Livin异构体基因沉默对肺腺癌SPC-A1细胞化疗增敏效应研究  

Study on enhancing sensitivity of SPC-A1 cells to chemotherapy by Livin isoform-specific gene silencing

作　　者：孙建国[1] 廖荣霞[2] 陈正堂[1] 王志新[1] 张青[3] 胡义德[1] 

机构地区：[1]第三军医大学新桥医院全军肿瘤研究所,重庆400037 [2]第三军医大学基础部生物化学与分子生物学教研室 [3]第三军医大学新桥医院核医学室,重庆400037

出　　处：《中国肺癌杂志》2007年第6期461-465,共5页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金青年项目(No30200282)资助~~

摘　　要：背景与目的前期研究发现,凋亡抑制蛋白家族(IAP)新成员Livin尤其是Livinα,参与肺癌的发生发展,是肺癌细胞化疗耐药的重要机制之一。本研究的目的是利用基因转染和RNA干涉(RNAi)技术,在肺腺癌SPC-A1细胞中建立Livin异构体α和β特异的基因沉默体系,探讨其对增加SPC-A1细胞化疗敏感性的不同作用及价值。方法在SPC-A1细胞中稳定表达Livinα+β、Livinα和Livinβ特异性小干涉RNA(siRNA),体外实验用甲基偶氮唑蓝(MTT)法,研究Livin基因沉默后SPC-A1细胞对化疗药物的敏感性改变;体内实验利用荷瘤裸鼠模型,研究Livin基因沉默后荷瘤小鼠对顺铂的敏感性改变。结果Livinα+β、Livinα和Livinβ基因沉默后,SPC-A1细胞对顺铂、卡铂、环磷酰胺、阿霉素等多种化疗药物敏感性增加(P<0.05),其中,Livinα+β基因沉默能更有效增加肺癌细胞化疗敏感性(P<0.01)。动物实验表明,pSilencer-Livinα+β组、pSilencer-Livinα组和pSilencer-Livinβ组抑瘤率分别为146.1%、130.7%、110.5%。结论Livin异构体尤其是Livinα+β可作为肺癌细胞化疗增敏的分子靶点,其基因沉默有望成为非小细胞肺癌基因治疗新手段。Background and objective As a new member of inhibitor of apoptosis protein(IAP) family,Livin,especially Livin α,is known to be involved in occurrence and development of lung cancer.Livin is an important mechanism of chemotherapy resistance of lung cancer cell.The aim of this study is to set up Livin isoform(α&β)-specific gene silencing system in SPC-A1 cells by gene transfection and RNA interference(RNAi),and to explore the different functions and value of the isoforms in enhancing chemosensitivity of SPC-A1 cells.Methods Livinα+β,Livinα and Livinβ specific siRNA were expressed stably in SPC-A1 cells,respectively.MTT was performed to study sensitivity of the cells to chemotherapy drugs.In vivo experiment was performed to test sensitivity of mouse bearing tumor to cisplatin after gene silencing of Livin.Results After silencing of Livinα+β,Livinα and Livinβ genes,sensitivity of SPC-A1 cells to many chemotherapy drugs(including cisplatin,carboplatin,cyclophosphamide and adriblastine) was markedly increased(P<0.05).Among them,gene silencing of Livinα+β showed the strongest enhancement effect on chemosensitivity of SPC-A1 cells(P<0.01).Animal experiment showed that tumor inhibition rate of pSilencer-Livinα+β,pSilencer-Livinα and pSilencer-Livinβ groups was 146.1%,130.7% and 110.5%,respectively.Conclusion The results suggest that Livin isoform,especially Livinα+β is hopeful to be a molecular target for increasing sensitivity of lung cancer cell to chemotherapy.Gene silencing may be a new means of gene therapy for non-small cell lung cancer.

关 键 词：凋亡抑制蛋白 LIVIN 凋亡 基因沉默 非小细胞肺癌 肿瘤耐药 

分 类 号：R734.2[医药卫生—肿瘤]