IFNα高表达对视黄酸诱导HL-60细胞增殖、分化的影响

Effects of retinoic acid on proliferation and differentiation in HL-60 cells wi th retroviral vector mediated IFNα gene transfer

目的 探讨IFNα基因转移与ATRA对HL 6 0细胞增殖抑制和诱导分化的协同作用。方法 选择对视黄酸类药物比较敏感的HL 6 0细胞株作为实验对象 ,以逆转录病毒载体pLXSN IFNα5经PA317细胞包装后感染HL 6 0细胞 ,RT PCR检测有IFNαmRNA表达 ,用ELISA检测IFNα分泌量 ,再以 2 .5× 10 - 7mol LATRA处理转染和未转染细胞 ,用流式细胞仪和NBT还原实验观察细胞的增殖、分化情况。结果 逆转录病毒载体介导IFNα在HL 6 0细胞中高表达 ,2 4h 10 6 个细胞的分泌量为95ng mL ,ATRA对IFNα高表达的HL 6 0细胞的增殖抑制及诱导分化作用显著增强。结论 通过逆转录病毒载体介导使IFNα在HL 6 0细胞中高表达 ,ATRA与HL 6 0细胞中高表达的IFNα具有协同作用。

ObjectiveTo study the synergistic eff ec ts of retinoic acid(RA) and IFNα gene transfer mediated by retroviral vector on proliferation and differentiation in HL 60 cells.MethodsHL 60 cells were infected with the supernatants of PA317 cells transfected with ret rovial vectors pLXSN IFNα. After analyzing the expression of IFNα mRNA by RT PCR and detectng the secretion of IFNα by ELISA, infected HL 60 cells we re treated with RA. For examining the changes of cellular proliferation capacity ,cel l cycle distribution and cellular differentiation,the flow cytometer analysis, N BT reduction test were applied.Results IFNα highly expressed i n HL 60 cells with retroviral vector me diated IFNα gene transfer. The pr olifer ation capacity decreased in cells treated by RA, the capability of reducing NBT increased in cells treated by RA. These changes were more obvious in transfected cells than non transfected cells.Conclusion Our study demonst rated that RA and IFNα gene transfer mediated by retroviral vector could cooper ate in inhibition of cell growth and inducing differentiation in HL 60 cells.