RNA干扰沉默缺氧诱导因子1α逆转肺癌细胞耐药性  

Reversing the Multidrug Resistance of Human Pulmonary Adenocarcinoma Cells by RNA Interference Targeting HIF-1alpha Gene

作　　者：刘胜岗[1] 陈琼[2] 何白梅[2] 陈莹[3] 聂华萍[1] 梁伟军[1] 

机构地区：[1]长沙市中心医院呼吸内科,湖南长沙410000 [2]中南大学湘雅医院老年医学科,湖南长沙410078 [3]湖南省人民医院肾内科,湖南长沙410005

出　　处：《现代生物医学进展》2011年第13期2486-2489,共4页Progress in Modern Biomedicine

摘　　要：目的:观察RNA干扰沉默缺氧诱导因子1α(HIF-1α)对肺癌细胞耐药性的影响。方法:构建靶向HIF-1α小干扰RNA基因,并转染到人肺腺癌耐顺铂细胞株A549/DDP细胞中。逆转录聚合酶链反应RT-PCR)检测细胞的HIF-1α、多药耐药基因1(MDR-1)以多药耐药相关蛋白基因(MRP)mRNA变化,免疫细胞化学法观察干扰后HIF-1α、P-糖蛋白以及MRP蛋白的变化。MTT法检测不同浓度的顺铂作用下细胞死亡率。结果:HIF-1αsiRNA组中HIF-1α、MDR-1、MRP mRNA水平显著降低(P<0.05),且蛋白水平也显著下降(P<0.05)。HIF-1αsiRNA组细胞死亡率较未转染组均明显增高(P<0.05),转染siRNA阴性组不影响肿瘤细胞的耐药性。结论:HIF-1αsiRNA可显著降低A549/DDP细胞中HIF-1α、MDR-1、MRP表达,从而起到逆转肺腺癌A549/DDP细胞的耐药作用。Objective： To reverse the multidrug resistant （MDR） phenotype of human lung cancer cells by small interference RNA （siRNA） technique targeting hypoxia-inducible factor-1α （HIF-1α） gene. Methods： Small interference RNA eukaryotic expression vector targeting HIF-1αgene was constructed and transfected into human adenocarcinoma A549/DDP cells by liposome technique. The mRNA and protein expression levels of HIF-1α, MDR-1 and MRP were investigated by RT-PCR and immuocytochemistry. The mortality of cells was observed by MTT in different groups treated with cisplatin of different concentrations. Results： mRNA and protein levels of HIF-1α, MDR-1 and MRP were significantly decreased in HIF-1α siRNA group. The MTT assays showed the mortality in HIF-1α siRNA group was significantly higher than that in non-transfected group （P 〈0.05）. The negative siRNA transfection did not affect the resistance of tumor cells. Conclusion： HIF-1α siRNA significantly reduced the expression ofHIF-1α, MDR-1 and MRP in A549/DDP cells and thus played a role in reversing drug resistance.

关 键 词：小RNA干扰 低氧诱导因子-1Α 多药耐药基因1 多药耐药相关蛋白 

分 类 号：R562[医药卫生—呼吸系统]