吡格列酮对K562/ADR细胞多药耐药的逆转作用及机制

Reversal Effect of Pioglitazone on Multidrug Resistance in K562/ADR Cells and Its Mechanism

目的:探讨吡格列酮对K562/ADR细胞阿霉素耐药的逆转作用及机制。方法:应用MTT法检测吡格列酮对K562及K562/ADR细胞的增殖抑制率、半数抑制浓度(IC50)、耐药逆转倍数并绘制增殖抑制率曲线;应用RT-PCR法测定PPARγ、CYP2C8和CYP2J2基因的转录;Western blot法测定PPARγ、CYP2C8和CYP2J2蛋白的表达。结果:吡格列酮作用于K562及K562/ADR细胞60 h的IC50分别为326μmol/L、349μmol/L;30μmol/L吡格列酮逆转K562/ADR细胞的阿霉素耐药倍数为6.4倍。吡格列酮作用于K562/ADR细胞后,CYP2C8和CYP2J2基因转录明显下降,CYP2C8、CYP2J2蛋白表达明显降低,PPARγ转录和PPARγ蛋白表达无差异。结论:吡格列酮可逆转K562/ADR细胞对阿霉素的耐药,其机制与CYP2C8和CYP2J2蛋白下调密切相关。吡格列酮是一种有效的肿瘤多药耐药逆转剂。

Objective:To explore the reversal effect of pioglitazone(PIO)on multidrug resistance in K562/ADR cells and its mechanism.Methods:The proliferation inhibition rate,half inhibition concentration(IC50)and drug-resistance reversal multipe were detected and the curve of proliferation inhibition rate was drawn by MTT assay,the transcription of PPARy,CYP2C8 and CYP2J2 genes was detected by RT-PCR;the expression of PPARγ,CYP2C8 and CYP2J2 proteins was detected by Western blot.Results:The IC50 of PIO on K562 and K562/ADR cells for 60 h was 326.7μmol/L and 349.1μmol/L respectively.The reversal multiple of 30μmol/L PIO on ADR-resistance of K562/ADR cells was6.4.After treatment of K562/ADR cells with PIO,the transcription of CYP2C8 and CYP2J2 and the protein expression of CYP2C8 and CYP2J2 significantly decreased,the transcription of PPARγgene and the expression of PPARγprotein were not changed.Conclusions:Pioglitazone can reverse the adriamycin-resistance in K562/ADR cells that is closely related to the decrease of protein expression of CYP2C8 and CYP2J2.Pioglitazone is an effective multidrug resistance reversal agent for tumors.