SIRT1-shRNA慢病毒表达载体转染对A549细胞生物学行为的影响  

Effect of SIRT1-shRNA lentiviral expression vector transfection on biological behavior of non-small-cell lung cancer A549 cell line

作　　者：孙硕文[1,2] 朱之燕[1] 杜玮[1] 王豪[1] 刘喆[1] 

机构地区：[1]天津医科大学,天津300070 [2]天津市胸科医院

出　　处：《山东医药》2015年第22期12-15,共4页Shandong Medical Journal

基　　金：天津市高等学校科技发展基金计划项目(20140602)

摘　　要：目的探讨SIRT1-shRNA慢病毒表达载体转染对人非小细胞肺癌A549细胞SIRT1基因表达、黏附能力、迁移能力及E-cadherin、β-catenin mRNA和蛋白表达量的影响。方法构建SIRT1-shRNA慢病毒表达载体,并用其转染A549细胞。对照组转染277空载体,观察组转染277-i SIRT1。用RT-PCR法检测两组细胞SIRT1 mRNA表达,用细胞黏附实验检测两组细胞黏附能力,划痕实验检测细胞迁移能力,RT-PCR法检测细胞内E-cadherin、β-catenin mRNA表达,Western blot法检测细胞内E-cadherin、β-catenin蛋白表达。结果观察组、对照组A549细胞SIRT1 mRNA分别为0.48±0.26、1.00±0.00(P<0.05),黏附细胞数分别为(196.6±9.8)、(125.6±9.8)个(P<0.05),细胞相对迁移率分别为65%±2%、100%±0(P<0.05),E-cadherin蛋白表达量分别为1.27±0.16、1.00±0.00(P<0.05),β-catenin蛋白表达量分别为1.24±0.13、1.00±0.00(P<0.05)。结论 SIRT1-shRNA慢病毒表达载体可明显下调A549细胞SIRT1表达,提高其下游基因E-cadherin、β-catenin mRNA和蛋白表达量,增强细胞黏附能力,降低细胞迁移能力。Objective To investigate effect of SIRT1-shRNA lentiviral expression vector transfection on the cell adhesion,migration and the expression of E-cadherin and β-catenin in non-small-cell lung cancer（ NSCLC） A549 cell line.Methods The SIRT1-shRNA lentiviral expression vector was constructed and we used it to transfect the A549 cells. The cells infected with 277 empty vector were used as the control group and the cells infected with 277-i SIRT1 were used as the observation group. The SIRT1 mRNA expression in the two groups was detected by RT-PCR. Cell adhesion status of A549 cells in the two groups was detected by cell adhesion assay,cell migration was detected by wound healing assay,and the protein and mRNA expression levels of E-cadherin andβ-catenin were detected by RT-PCR and Western blotting,respectively. Results the SIRT1 mRNA of A549 cells in the observation group and control group were respectively 0. 48 ± 0. 26 and 1. 00 ± 0. 00（ P〈0. 05）,the adhesive cells of the observation group and control group were 196. 6 ± 9. 8 and 125. 6± 9. 8,respectively（ P〈0. 05）,the relative migration rates of the observation group and control group were 0. 65 ± 0. 02 and 1. 00 ± 0. 00,respectively（ P〈0. 05）. and the E-cadherin expression levels of the observation group and control group were 1. 27 ± 0. 16 and 1. 00 ± 0. 00（ P〈0. 05）,and β-catenin expression levels were 1. 24 ± 0. 13 and 1. 00 ± 0. 00（ P〈0. 05）. Conclusion SIRT1-shRNA lentiviral expression vector may significantly down-regulate the expression of SIRT1 in A549 cells,increase the expression levels of E-cadherin andβ-catenin mRNA and protein,enhance the capacity of cell adhesion and reduce the cell migration ability.

关 键 词：非小细胞肺癌 SIRT1-shRNA慢病毒表达载体 SIRT1基因 E-CADHERIN蛋白 Β-CATENIN蛋白 细胞黏附 细胞迁移 

分 类 号：R734.2[医药卫生—肿瘤]