摘要
目的 探讨视黄酸 (retinoicacid ,RA)及其诱导的反义 (anti sense,AS)cyclinD1对HL 6 0细胞的细胞周期素激酶CDK4的影响。方法 通过构建含有视黄酸反应元件 (retinoicacidresponseelement,RARE)的反义cyclinD1RNA表达载体 pCI neo/RARE3 TK/AscyclinD1,使反义cyclinD1的表达可受视黄酸诱导调控。以脂质体转染HL 6 0白血病细胞后 ,运用MTT比色法检测细胞增殖功能、NBT还原实验观测细胞分化状况 ,RT PCR以及western blot等方法观测视黄酸处理后CDK4的mRNA和蛋白表达水平的改变。结果 RA处理后 ,细胞生长显著减慢 ,分化能力明显增强 ,CDK4mRNA和蛋白表达水平均有所降低 ,其中 ,以反义cyclinD1转染细胞经RA处理后变化更为显著。结论 RA及其诱导的反义cyclinD1对HL 6
Objective To study the effect for anti-sense cyclin D1 dependent on retinoic acid(RA) on the cyclin dependent kinase 4 (CDK4) of HL-60 cells. Methods After transfection of anti-sense cyclin D1 RNA expression vector (pCI-neo/RARE3-TK/Ascyclin D1)and treatment of RA in HL-60 cells, MTT was used for cell proliferation analysis, nitroblue tetragolium reduction experiment for cell differentiation, reverse transcriptase polymerase chain reaction(RT-PCR) for CDK4 mRNA, and western-blot for CDK4 protein expression. Results The proliferation capacity was decreased, the differentiation capacity was increased and the expression of CDK4 mRNA and protein were decreased in cells by RA. These changes were more obviously in transfected cells than nontransfected cells. Conclusion RA and the product of anti-sense cyclin D1 induced by RA could cooperate in resisting cell proliferation and inducing cell differentiation, the down-regulated expression of CDK4 may play an important role in the molecular mechanism.
出处
《肿瘤》
CAS
CSCD
北大核心
2004年第4期329-332,共4页
Tumor
基金
国家自然科学基金资助项目 (3 0 0 70 3 2 8)
