摘要
目的 探讨全反式维甲酸诱导分化的HL6 0细胞周期变化 ,进一步揭示细胞分化在细胞周期中的时相特异性。方法 以分化诱导剂全反式维甲酸 (终浓度为 10 μmol/L)影响下不同时间点的HL6 0细胞为检测对象 ,应用流式细胞术分析细胞的大小及细胞表面的分化标志 ;碘化丙啶 (PI)染色后用激光共聚焦显微镜观察细胞形态从而对已分化细胞进行确认 ;应用流式细胞术分析药物诱导的细胞周期变化 ,检测G1期的Ki6 7表达水平 ;再应用流式分选术结合激光共聚焦显微镜观察各期细胞形态。结果 ①随着药物诱导时间的延长 ,细胞的体积逐渐增大 ,被诱导的细胞出现髓系细胞表面的分化标志物CD11b。②全反式维甲酸导致HL6 0细胞周期的G0 /G1峰升高 ,S期水平下降。③随着药物诱导时间的延长 ,G1期的Ki6 7的表达水平逐渐下降。④只在G1期细胞中可见到已分化细胞。结论 全反式维甲酸可以诱导HL6 0细胞周期的G1早期→G1晚期阻滞 ,并诱导HL6 0沿着粒系方向分化 ,细胞分化完成于细胞周期中的G1早期。
Objective To investigate the changes in the cell cycle of differentiated HL60 cells initiated by all trans retinoic acid (ATRA) and explore the specific phase of the cell differentiation in cell cycle. Methods HL60 cells stimulated by ATRA (10 μmol/L) capable of inducing differentiation for different time were used as subjects. The cell volume and the differentiation marker on the cell membrane surface were analyzed by using flow cytometry. The differentiated cells were identified by confocal microscope after having been stained with propidium iodide (PI). Meanwhile, the change of the cell cycles induced by those drugs was analyzed by using flow cytometry. The expression level of Ki67 in G 1 phase was detected by using flow cytometry. The specific phase of the drug-induced cell differentiation in cell cycle was explored by flow cytometry sorting combined with confocal microscopic pictures of the cells in different phases of cell cycle. Results When the drug-induced time prolonged, the induced cell volume enlarged and the induced cells began to express CD11b (differentiation marker of myeloid cell). The peak value of G 0/G 1 phase was elevated in drug-induced cell cycle and the level of S phase was declined in drug-induced cell cycle. The expression level of Ki67 in G 1 phase was gradually declined. The differentiated cell was only observed in G 0/G 1 phase. Conclusion ATRA can induce HL60 cell to differentiate along the way to granulocytic cells and induce the arrest of cell cycle in early G 1 phase to late G 1 phase. The cell differentiation was completed in early G 1 phase in cell cycle.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2004年第4期445-448,514,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金(No.39730270)
国家杰出青年基金(No.39725027)
973肿瘤计划(No.G1998051212)
卫生部临床重点学科基金(No.20012537)资助项目
