脑胶质瘤相关癌基因1对人胰腺癌PANC-1细胞增殖的影响  

Effect of glioma-associated oncogene 1 on the proliferation of human pancreatic cancer PANC-1 cells

作　　者：王锋[1] 徐凌[1] 郭传勇[1] 杨丽娟[1] 何姗姗[1] 徐选福[1] 黄银实[1] 卫巍[1] 戴维奇[1] 沈杰[1] 王兴鹏[1] 

机构地区：[1]同济大学附属第十人民医院消化科,上海200072

出　　处：《肿瘤》2011年第5期400-405,共6页Tumor

基　　金：国家自然科学基金资助项目(编号:81072005);上海市科学技术委员会科研计划项目(编号:08411963000);上海市卫生局科研课题(编号:2009142)

摘　　要：目的:探讨脑胶质瘤相关癌基因1(glioma-associated oncogene1,GLI1)对人胰腺癌PANC-1细胞增殖的影响。方法:构建靶向GLI1基因的RNA干扰(RNAinterference,RNAi)慢病毒重组载体pLVTHM-GLI1-shRNA(以空载体pLVTHM-GFP为对照),经包装后感染胰腺癌PANC-1细胞,应用实时荧光定量PCR(real-time fluorescence quantitative-PCR,RFQ-PCR)和蛋白质印迹法检测各组细胞中GLI1、bcl-2、bcl-xl和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)mRNA和蛋白的表达;CCK8(cell counting kit-8)法分析各组细胞的生长情况。结果:成功构建了稳定低表达GLI1的GLI1-shRNA-PANC-1细胞,与PANC-1细胞和GFP-PANC-1细胞比较,GLI1-shRNA-PANC-1细胞中GLI1mRNA和蛋白表达水平分别下调了(82.1±3.2)和(76.7±2.2)(P<0.01),bcl-2mRNA和蛋白表达分别下调了(49.7±5.4)、(43.5±9.4)(P<0.01);而blc-xl和PCNAmRNA和蛋白表达水平无明显变化。GLI1-shRNA-PANC-1细胞的增殖抑制率明显高于GFP-PANC-1、PANC-1细胞(P<0.05)。结论:干扰GLI1基因的表达可抑制人胰腺癌PANC-1细胞的增殖,其机制可能与下调bcl-2的表达有关。Objective：To investigate the effect of glioma-associated oncogene 1（GLI1） on the proliferation ability of human pancreatic cancer PANC-1 cells.Methods：The recombinant lentiviral vector of RNA interference（RNAi）-targeting GLI1 gene was constructed.The pLVTHM-GLI1-shRNA or pLVTHM-GFP lentivirus（as the control）was infected into PANC-1 cells.The expressions of GLI1,bcl-2,bcl-xl and proliferating cell nuclear antigen（PCNA） mRNAs and proteins were detected by real-time fluorescence quantitative-PCR and Western blotting,respectively,and the cell growth was measured by cell count kit-8（CCK8） assay.Results：The GLI1-shRNA-PANC-1 cells with stable low-expression of GLI1 were successfully established.Compared with GFP-PANC-1 and PANC-1 cells,the expression levels of GLI1 mRNA and protein in GLI1-shRNA-PANC-1 cells were down-regulated by（82.1±3.2）% and（76.7±2.2）%,respectively（P0.01）,and the expression levels of bcl-2 mRNA and protein were down-regulated（49.7±5.4）% and by（43.5±9.4）%,respectively（P0.01）.There were no obvious changes in expressions of bcl-xl or PCNA mRNA and protein among three groups.The proliferation inhibitory rate of GLI1-shRNA-PANC-1 cells was higher than those in GFP-PANC-1 and PANC-1 cells（P0.05）.Conclusion：GLI1 RNAi can inhibit the proliferation ability of pancreatic cancer PANC-1 cells.This effect may be associated with the down-regulation of bcl-2 expression.

关 键 词：胰腺肿瘤 RNA干扰 细胞增殖 慢病毒属 脑胶质瘤相关癌基因1 细胞 PANC-1 

分 类 号：R735.9[医药卫生—肿瘤]