HIF—1α对人肝癌细胞MDR1表达的影响

THE EFFECT OF HIF -1α ON MULTI -DRUG RESISTANCE IN HUMAN HEPATOCARCINOMA CELLS

目的探讨缺氧诱导因子-1(hypoxia-inducible factors,HIF-1)α对人肝癌细胞株多药耐药基因(multi-drug resistance gene 1,MDR1)mRNA和蛋白表达的影响。方法以0、1.0、3.0、5.0μmol/L浓度的HIF-1α抑制剂YC-1和0、1.0、2.5、5.0μmol/L HIF-1α的反义寡核苷酸作用于人肝癌细胞株BEL-7402细胞,24h后RT-PCR法检测药物干预前后肝癌细胞株HIF-1、MDR1mRNA的变化;Western-Blot方法检测肝癌细胞株HIF-1、MDR1蛋白表达的变化。结果①反转录-聚合酶链反应产物结果显示,YC-1作用后,肝癌细胞HIF-1αmRNA表达差异无统计学意义,随着HIF-1α抑制剂浓度的增加,MDR1 mRNA的表达逐渐降低;随着HIF-1α反义寡核苷酸浓度的增加,HIF-1αnRNA和MDR1mRNA的表达逐渐减少,差异有统计学意义(P<0.05)。②Western-Blot检测结果显示,随着HIF-1α抑制剂浓度增加,BEL-7402细胞HIF-1α、MDR1蛋白表达逐渐减少;随着HIF-1α反义寡核苷酸浓度增加,BEL-7402细胞HIF-1α、MDR1蛋白表达亦逐渐减少。药物干预组与对照组、各浓度组比较差异有统计学意义(P<0.05)。结论 HIF-1α可以促进肝癌细胞MDR1mRNA和蛋白的表达。

Objective To investigate the regulation and influence of hypoxia inducible factor - 1 α （ HIF - 1 ） on the gene expression of multi - drug resistance genel （ MDR1 ） in human hepatocarcinoma cell lines. Methods BEL- 7402 cells were divided into control group and experiment group. The former were cultured with normal condition, the later were treated by YC - 1 , an inhibitor of HIF - 1, or the antisense oligodeoxynucleotides of HIF - 1 （ AS - ODN） with various concentration, and all of the cells were collected on time. The mRNA expressions of HIF - 1 and MDR1 in the cultured BEL -7402 cells were evaluated by reverse transcription - polymerase chain reaction and the protein levels of HIF - 1 and MDR1 in cytoplasm were evaluated by western blot. Results In human hepatocarcinoma cells, with the increasing of the concentration of YC - 1 or AS - ODN, mRNA expressions of HIF - 1 and MDR1 were decreased and the protein expressions of HIF - 1 and MDR1 were also decreased . There were significantly differences between the control and experiment groups （ P 〈 0.05 ）. Conclusion HIF - 1 can promote the expression of MDR1 at the mRNA and protein levels in human hepatocarcinoma cells.