视黄酸依赖反义cyclin D1对HL-60细胞分化效应研究

目的 检测视黄酸(Retinoic acid,RA)依赖反义cyclin D1表达载体转染HL-60细胞后启动的诱导分化效应,并探讨其分子机制。方法 成功构建视黄酸依赖反义cyclin D1表达载体,以脂质体转染HL-60白血病细胞,经NBT还原实验、免疫荧光检测CD14表面抗原表达、RT-PCR以及Western blot等方法观测视黄酸处理后细胞分化效应的发生以及Rb基因的mRNA和蛋白表达水平的改变。结果 反义eyelin D1转染和未转染的HL-60细胞经RA处理后,NBT还原能力明显增强、CD14表面抗原表达显著增加,Rb基因的mRNA和蛋白表达水平均有所降低,其中,以反义cyclin D1转染细胞经RA处理后变化更为明显。结论 RA及其诱导的反义cyclin D1可协同诱导HL-60细胞分化成熟,下调Rb基因表达可能是其分子机制之一。

Effects of anti-sense cyclin D1 induction in retinoic acid dependent manner on differentiation of HL-60 cells

Objective To study the effect of anti-sense cyclin D1 induction in retinoic acid dependent manner on the differentiation of HL-60 cells and its mechanism. Methods After transfection of anti-sense cyclin D1 RNA expression vector and treatment of retinoic acid (RA) in HL-60 cells, NBT reduction experiment was used for cell differentiation, immunofluorescence for the expression of CD14 surface antigen, reverse transcriptase polymerase chain reaction(RT-PCR) for retinoblastoma(Rb) mRNA, and western-blot for Rb protein(pRb) expression. Results The NBT reduction capacity and the expression of CD14 surface antigen were increased in cells by RA. The expression of Rb mRNA and pRb were decreased in cells by RA. These changes were more obvious in transfected cells than non-transfected cells. Conclusion RA and the product of anti-sense cyclin D1 induced by RA could cooperate in inducing cell differentiation, the down-regulated expression of Rb may play an important role in the molecular mechanism.