文章摘要
刘胜岗1 陈琼2△ 何白梅2 陈莹3 聂华萍1 梁伟军1.RNA 干扰沉默缺氧诱导因子1α逆转肺癌细胞耐药性[J].,2011,11(13):2486-2489
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 AdenocarcinomaCells by RNA Interference Targeting HIF-1alpha Gene
  
DOI:
中文关键词: 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
英文关键词: Small interference RNA  Hypoxia-inducible factor-1α  Multidrug-resistance-1  Multidrug resistance-associated protein
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刘胜岗1 陈琼2△ 何白梅2 陈莹3 聂华萍1 梁伟军1 长沙市中心医院呼吸内科 
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中文摘要:
      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-1α siRNA 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。
英文摘要:
      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 of HIF-1α, MDR-1 and MRP in A549/DDP cells and thus played a role in reversing drug resistance.
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