hp450RAI质粒转染对视黄酸诱导的HL-60细胞凋亡的影响

Effects of human cytochrome p450 enzyme gene (hp450RAI) transfer into HL-60 cells on apoptosis induced by retinoic acid

目的探讨细胞内细胞色素氧化酶 p45 0 (CYP)表达、视黄酸 (RA)代谢与细胞凋亡的相互关系。方法采用脂质体介导的质粒转染和逆转录 - PCR等实验技术方法做有关分析检测。结果野生型 HL- 6 0细胞 hp45 0 RAI表达微弱 ,脂质体介导的 hp45 0 RAI质粒转染可使 HL- 6 0细胞稳定表达 hp45 0 RAI;hp45 0 RAI转染对 ATRA诱导的 HL- 6 0细胞增殖能力无明显影响 ;野生型 HL- 6 0细胞有较强 Bcl- 2表达 ,ATRA处理抑制未转染细胞 Bcl- 2表达 ,但单纯 hp45 0 RAI转染、hp45 0 RAI转染 +CYP抑制剂或加用 IFN- α均能增强 Bcl- 2表达 ,其中以加用 ketoconazole作用更为显著。结论 RA敏感 HL- 6 0细胞中影响 RA代谢的主要 CYP并非 hp45 0 RAI,hp45 0 RAI转染能增强 Bcl- 2表达 ,但对 HL - 6 0细胞增殖能力无明显影响 ,CYP抑制剂、IFN-α可能通过抑制细胞内 RA代谢 ,而增加 HL - 6 0细胞Βcl- 2表达。

Objective[WT5”BZ]To investigate the relationship of human cytochrome p450 enzyme gene(hp450RAI) expression,retinoic acid metabolism and cell apoptosis in HL 60 cell.Methods The plasmid transfer mediated by liposomal transfectin reagent and reverse transcription PCR analysis were used in this study.ResultsThe hp450RAI mRNA was weakly detected in wild HL 60 cells,but the gene transfer made the HL 60 cells express hp450RAI greatly.The hp450RAI gene transfer could not affect the growth of HL 60 cells induced by ATRA,Bcl 2 gene was expressed obviously in HL 60 cells,ATRA treatment could inhibit its expression,but hp450RAI transfer,and combined with the ketoconazole or IFN α increased the Bcl 2 mRNA in ATRA treated HL 60 cells.Conclusion hp450RAI may not be the main cytochrome p450 enzyme which modulates the metabolism of retinoic acid in wild HL 60 cells.hp450RAI transfer into HL 60 cells can not affect the cellular proliferation,but Bcl 2 expression can be changed by the hp450RAI transfer,cytochrome enzyme inhibitor or IFN α in HL 60 cells. [