GRIM-19基因对肺腺癌耐药细胞株A549/DDP药物敏感性及相关基因表达的影响  

Effects of GRIM-19 on chemotherapeutic sensitivity of cisplatin-resistant human lung cancer cell A549/DDP and the expressions of related genes

作　　者：武道荣[1,2] 王同[3] 江子丰[1,2] 刘荣玉[1,2] 

机构地区：[1]安徽医科大学第一附属医院干部呼吸内科,合肥230022 [2]安徽医科大学呼吸病研究所,合肥230022 [3]合肥市第一人民医院呼吸内科,合肥230061

出　　处：《肿瘤》2012年第2期92-98,共7页Tumor

基　　金：安徽省科技计划资助项目(编号:10021303028)

摘　　要：目的:本研究旨在探讨干扰素/维甲酸诱导凋亡相关基因19(gene-associated with retinoid-interferon-induced mortality19,GRIM-19)对人肺腺癌耐药细胞株A549/DDP化疗药物敏感度及相关基因表达的影响。方法:采用脂质体法将重组质粒PIRES-Puro2-GRIM-19-Myc和空载体PIRES-Puro2-Myc分别转染顺铂(cisplatin,DDP)耐药的人肺腺癌细胞株A549/DDP。蛋白免疫印迹法检测亲本A549、耐药A549/DDP、转染GRIM-19基因的A549/DDP及转染空载体的A549/DDP细胞中GRIM-19蛋白的表达。MTT法检测稳定转染GRIM-19基因后A549/DDP细胞对多种化疗药物敏感度的改变。实时荧光定量-PCR(real-time fluorescence quantitative-PCR,RFQ-PCR)法检测GRIM-19、信号转导子与转录激活子3(signal transducers and activators of transcription 3,STAT3)、血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)和P-糖蛋白(P-glycoprotein,P-gp)mRNA在稳定转染GRIM-19基因后A549/DDP细胞中的表达情况。结果:蛋白质印迹法检测结果显示,A549/DDP细胞转染GRIM-19基因后GRIM-19蛋白的表达上调;A549/DDP细胞较亲本A549细胞对DDP的耐药指数为16.86±1.32;A549/DDP细胞转染GRIM-19基因后,较转染空载体组A549/DDP细胞耐药性降低,耐药逆转倍数为3.70±0.91。转染GRIM-19基因后,A549/DDP细胞中STAT3、P-gp和VEGFmRNA表达均下调。结论:GRIM-19能增强DDP耐药细胞A549/DDP对化疗药物的敏感度,这可能与下调STAT3、VEGF和P-gp的表达具有相关性,GRIM-19可能成为逆转A549/DDP细胞耐药性的一条有效途径。Objective：To investigate the effects of gene-associated with retinoid-interferon-induced mortality 19（GRIM-19） on chemotherapeutic sensitivity of cisplatin（DDP）-resistant human lung caner cell A549/DDP and the expressions of related genes.Methods：The recombinant plasmid PIRES-Puro2-GRIM-19-Myc and the empty vector PIRES-Puro2-Myc were transfected into A549/DDP cells by liposome transfection reagent,respectively.The expression levels of GRIM-19 protein in the A459 parental cells,A459/DDP cells,A549/DDP cells transfected with GRIM-19 gene and A549/DDP cells transfected with an empty vector were detected by Western blotting.The changes of chemotherapeutic sensitivities to multi-drugs in A549/DDP cells stably transfected with GRIM-19 gene were detected by MTT assay.The expression levels of GRIM-19,signal transducers and activators of transcription 3（STAT3）,vascular endothelial growth factor（VEGF） and P-glycoprotein（P-gp） mRNAs in the A549/DDP cells stably transfected with GRIM-19 gene were examined by real-time fluorescence quantitative-PCR（RFQ-PCR）.Results：The Western blotting result showed that the expression level of GRIM-19 protein was increased in the A549/DDP cells after transfection with GRIM-19 gene.The resistance index of A549/DDP cells was 16.86±1.32 folds higher than that of the A549 parental cells.The resistance index of the A549/DDP cells after transfection with GRIM-19 gene was decreased by 3.70±0.91 folds as compared with that of the A549/DDP cells transfected with an empty vector.The expression levels of STAT3,VEGF and P-gp mRNAs in the A549/DDP cells transfected with GRIM-19 gene were decreased.Conclusion：GRIM-19 can increase the chemotherapeutic sensitivity of A549/DDP cells.This effect may be associated with the down-regulations of STAT3,VEGF and P-gp.GRIM-19 may become a potential therapeutic agent to reverse drug-resistance in A549/DDP cells.

关 键 词：癌 非小细胞肺 干扰素/维甲酸诱导凋亡相关基因-19 药物耐受性 转染 A549/DDP细胞 

分 类 号：R734.2[医药卫生—肿瘤]