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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 observe the effect of RNAi silencing hypoxia inducible factor 1α (HIF-1α) on the drug resistance of lung cancer cells. Methods: The HIF-1α small interfering RNA gene was constructed and transfected into human lung adenocarcinoma resistant cisplatin cell line A549 / DDP cells. The expression of HIF-1α and multidrug resistance gene 1 (MDR-1) were detected by reverse transcription-polymerase chain reaction (RT-PCR) -1α, P-glycoprotein and MRP protein. MTT assay was used to detect the cell death rate under different concentrations of cisplatin. Results: The levels of HIF-1α, MDR-1 and MRP mRNA in HIF-1αsiRNA group were significantly decreased (P <0.05), and the protein levels were significantly decreased (P <0.05). The cell death rate of HIF-1αsiRNA group was significantly higher than that of untransfected group (P <0.05). The transfected siRNA negative group did not affect the tumor cell resistance. Conclusion: HIF-1α siRNA can significantly reduce the expression of HIF-1α, MDR-1 and MRP in A549 / DDP cells, which can reverse the drug resistance of A549 / DDP cells.