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目的探讨缺氧环境下人肝癌细胞中多药耐药相关基因和缺氧诱导因子1α(HIF1α)的表达和意义,从而部分阐明肝细胞癌发生多药耐药的机制,为逆转肝癌耐药提供新的分子靶点。方法将人肝癌细胞系HepG2细胞分别行不同时间低氧培养和转染HIF1α/PCDNA3质粒;应用荧光定量聚合酶链反应技术和蛋白免疫印迹技术分别检测每组HepG2细胞中多药耐药相关基因(mdr1)、多药耐药相关蛋白1(MRP1)和肺耐药相关蛋白(LRP)在mRNA和蛋白水平的表达。结果在缺氧组,随着缺氧时间的延长HepG2细胞中多药耐药相关基因mdr1、MRP1和LRP的表达均逐渐增高,且以MRP1变化更为显著;而且这些多药耐药相关基因的表达升高与缺氧诱导因子1α的表达呈同步化改变。在HIF1α/PCDNA3质粒转染细胞中这些多药耐药相关基因的表达亦明显升高。结论缺氧可通过核转录因子HIF1α上调肝癌细胞内mdr1,LRP、MRP1等多药耐药相关基因的表达,从而使肝细胞癌获得多药耐药性。生长局部微环境的缺氧是诱导肝癌产生多药耐药性的重要原因之一。核转录因子HIF1α和这些多药耐药相关基因将可能成为逆转肝癌耐药的新的分子靶点。
Objective To investigate the expression and significance of multidrug resistance-related genes and hypoxia-inducible factor-1α (HIF1α) in human hepatocarcinoma cells under hypoxia, so as to elucidate the mechanism of multidrug resistance in hepatocellular carcinoma and to reverse the drug resistance of hepatocellular carcinoma New molecular target. Methods Human hepatocellular carcinoma HepG2 cells were cultured in different times at different times and transfected with HIF1α / PCDNA3 plasmid respectively. Fluorescent quantitative polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression of multidrug resistance-related genes in HepG2 cells mdr1), multidrug resistance-associated protein 1 (MRP1) and lung resistance-related protein (LRP) at mRNA and protein levels. Results In hypoxia group, the expression of mdr1, MRP1 and LRP in HepG2 cells gradually increased with the increase of hypoxia time, and the change of MRP1 was more significant. Moreover, the expression of multidrug resistance-related genes Increased expression and hypoxia inducible factor 1α expression showed a synchronized change. The expression of these MDR-related genes was also significantly increased in HIF1α / PCDNA3 plasmid-transfected cells. Conclusion Hypoxia can up-regulate the expression of multidrug resistance-related genes mdr1, LRP and MRP1 in hepatocellular carcinoma cells by HIF1α, thereby obtaining multidrug resistance in hepatocellular carcinoma. Hypoxia is one of the important reasons for inducing multidrug resistance in HCC. The nuclear transcription factor HIF1α and these MDR-related genes may be new molecular targets for reversing the drug resistance of hepatocellular carcinoma.