视黄酸依赖反义cyclin D1对HL-60细胞增殖和分化的影响及其机制

目的 :探讨视黄酸 (retinoicacid ,RA)及其诱导的反义 (anti sense ,AS)cyclinD1对肿瘤细胞的协同抑制增殖与诱导分化效应及其机制。方法 :通过构建的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RA诱导调控。以脂质体转染HL 60白血病细胞后 ,运用生长曲线、MTT比色法和流式细胞仪分析检测细胞增殖功能 ,NBT还原实验和透射电镜观测细胞分化状况 ,western blot检测p16基因蛋白表达水平。结果 :反义cy clinD1转染和未转染的HL 60细胞经RA处理后 ,生长减慢 ,细胞被阻滞在G0 /G1期 ,分化能力明显增强 ,p16基因蛋白表达增加 ,其中 ,以反义cyclinD1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cyclinD1可协同抑制HL 60细胞增殖和诱导细胞分化成熟 ,上调p16基因表达可能是其分子机制之一。

Effects of anti-sense cyclin D1 inducing in retinoic acid dependent manner on the proliferation and differentiation of HL-60 cells and its mechanism

OBJECTIVE:To study the effect of anti sense cyclin D1 inducing in retinoic acid dependent manner on the proliferation and differentiation of HL 60 cells and its mechanism.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,growth curve,MTT and flow cytometer detection were used for cell proliferation analysis,NBT reduction experiment and transmission electron microscope observation for cell differentiation,and western blot for the protein expression of p16.RESULTS:The proliferation capacity was decreased,the differentiation capacity was increased and the protein expression of p16 was increased in cells by RA.These changes were more obvious in transfected cells than those in nontransfected cells.CONCLUSIONS:RA and the product of anti sense cyclin D1 induced by RA could cooperate in resisting cell proliferation and inducing cell differentiation,and the up regulated expression of p16 may play an important role in the molecular mechanism.