hp450RAI质粒转染对HL-60细胞功能及IFN-α表达的影响

Effects of hp450RAI plasmid transfection on cellular function and IFN-α expression of HL-60 cells

目的探讨细胞内一种细胞色素 P45 0 (hp45 0 RAI,也称 CYP2 6 )表达、视黄酸 (RA)代谢与细胞增殖功能的相互关系。方法采用脂质体介导的质粒转染、流式细胞仪分析、逆转录 - PCR及高效液相色谱 (HPL C)分析等实验技术方法做有关分析研究。结果 12 .5× 10 - 7m ol/ L 全反式 RA(ATRA)处理 HL- 6 0细胞 3d未诱导明显的 hp45 0 RAI表达 ,脂质体介导的质粒转染可使 HL- 6 0细胞稳定表达 hp45 0 RAI;2单纯 hp45 0 RAI转染未对 HL- 6 0细胞 ATRA代谢及 ATRA诱导的 HL- 6 0细胞增殖抑制产生显著影响 ,而联合应用 P45 0酶抑制剂 Ketoconazole或 IFN- α则使 HL- 6 0细胞 ATRA代谢减慢 ,ATRA增殖抑制作用增强。结论 RA敏感 HL- 6 0细胞中影响 RA代谢的主要细胞色素 P45 0可能并非 hp45 0 RAI,P45 0酶抑制剂或IFN- α能协同 ATRA抑制细胞增殖 ,后者可能与 HL- 6 0细胞 ATRA代谢减慢及 IFN- α表达增强有关。

ObjectiveTo investigate the effects of human cytochrome P450 enzyme gene(hp450RAI)transfer on retinoic acid metabolism and cellular proliferation in HL-60 cells.Methods The plasmid transfection mediated by liposomal transfection reagent, HPLC analysis and reverse transcription-PCR analysis were used in this study. ResultsIt was shown that the hp450RAI mRNA was not induced by the treatment of 2.5×10 -7 mol/L all-trans retinoic acid(ATRA) in HL-60 cells; The hp450RAI gene transfer made the HL-60 cells express hp450RAI greatly, which did not affect significantly the cell growth and metabolism of ATRA in HL-60 cells, but the use of Ketoconazole or IFN-α could cut down the metabolic rate of ATRA in HL-60 cells,and enhance the growth inhibitory induced by ATRA. ConclusionThe hp450RAI may not be the key cytochrome P450 enzyme to modulate the metabolism of retinoic acid in wild HL-60 cells. The hp450RAI inhibitor ketoconazole or IFN-α can cooperate with ATRA to inhibit the growth of HL-60 cells, which may partly owe to the low metabolism rate of ATRA and over-expression of IFN-α in HL-60 cells. [