Hypoxia-induced increases in A549/CDDP cell drug resistance are reversed by RNA interference of HIF-1α expression
Hypoxia-induced increases in A549/CDDP cell drug resistance are reversed by RNA interference of HIF-1α expression
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DOI：
10.3892/mmr.2011.604
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发表时间：
2012-01-01
影响因子：
3.4
通讯作者：
Ma, Yun
中科院分区：
文献类型：
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作者：
Min, Lingfeng;Chen, Qiong;Ma, Yun
本研究旨在通过RNA干扰下调低氧诱导因子-1α(HIF-1α)对缺氧诱导的A549/CDDP细胞耐药的影响，并探讨其机制。构建了针对HIF-1α的小干扰RNA(SiRNA)真核表达载体，并将其导入缺氧处理的A549/CDDP细胞。用逆转录聚合酶链式反应(RT-PCR)和免疫细胞化学方法检测HIF-1α、多药耐药相关蛋白(MRP)和多药耐药相关蛋白(MDR1)的表达。用四甲基偶氮唑盐比色法测定顺铂作用后的细胞存活率。低氧增加了A549/CDDP细胞对顺铂的耐药性，这种作用可被siRNA抑制HIF-1α的表达所逆转。HIF-1αsiRNA的表达也下调了缺氧处理的A549/CDDP细胞中HIF-1α、mdr1和MRP的mRNA和蛋白表达(p
This study aimed to investigate the effects of knocking down hypoxia-inducible factor-1 alpha (HIF-1 alpha) through RNA interference on hypoxia-induced increases in drug resistance in A549/CDDP cells, and to study the underlying mechanisms. A small interfering RNA (siRNA) eukaryotic expression vector targeting HIF-1 alpha was constructed and transfected into A549/CDDP cells treated with hypoxia. The mRNA and protein levels of HIF-1 alpha, multidrug resistance-1 (MDR1), and multidrug resistance-associated protein (MRP) were determined by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry. Cell viability following treatment with cisplatin was determined by MTT assay. Hypoxia increased the resistance of A549/CDDP cells to cisplatin and this effect was reversed by the siRNA inhibition of HIF-1 alpha expression. Expression of HIF-1 alpha siRNA also downregulated HIF-1 alpha, MDR1 and MRP mRNA, and protein expression in A549/CDDP cells treated with hypoxia (p