视黄酸和α-干扰素对带有视黄酸反应元件的LUC报告基因表达的调控作用

Effect of retinoic acid on the expression of luciferase reporter gene in HL-60 cells transfected with the reporter plasmids containing retinoic acid response element

目的 探讨视黄酸 (Retinoicacid ,RA)通过视黄酸反应元件 (RARE)调控外源基因表达的可行性。方法 构建带有视黄酸受体 β(RARβ)基因增强子区域RARE的荧光素酶 (Luciferase ,LUC)报告基因表达载体 ,应用脂质体包裹RARE TK LUC和RARE3 TK LUC质粒 ,并转染HL 60细胞 ,48h后 ,用RA和 /或α 干扰素 (IFNα)处理不同时间 ,用LUC报告基因检测试剂盒分析LUC报告基因活性。结果 经RA处理不同时间后 ,RARE TK LUC和RARE3 TK LUC质粒转染细胞的LUC相对活性分别较未经处理的转染细胞增加 12～ 3 8和 2 0～ 85倍 ;单用IFNα处理转染细胞 ,对LUC的表达无明显的诱导作用 ;IFNα和RA联合处理 48、72h ,LUC相对活性比单用RA处理增加 60 %～ 70 %。结论 RA可调控带有RARE的LUC报告基因的表达 。

Objective To investigate the possibility of the expression of exogenous gene mediated by retinoic acid(RA). Methods Two reporter plasmids containing retinoic acid response element(RARE) ,pRARE TK LUC, and pRARE3 TK LUC, were transfected into HL 60 cells with liposome DOTAP respectively. After a period of 48 h, luciferase activity of transfected cells stimulated by vehicle or all trans retinoic acid(ATRA) and/or interferon alpha(IFNα) was determined by measurement of relative luciferase units in scintillation counter device. Results Stimulations of transient transfected HL 60 cells with ATRA within differential scheduled periods resulted in 12～38 fold and 20～85 fold increase in luciferase activity over basal activity of pRARE TK LUC and pRARE3 TK LUC transfected cells respectively. A combination of RA and IFNα led to a higher increase in luciferase activity than that merely by RA or IFNα. Conclusion Retinoic acid could induce the expression of exogenous gene containing RARE.