| 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 |
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| 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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| 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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