六亚甲基二乙酰胺诱导HL-60细胞分化及其分子机制的研究  

Molecular Mechanism Underlying Differentiation of HL-60 Cells Induced by Hexamethylene Bisacetamid

作　　者：任霞[1] 温培娥[1] 杨炜华[1] 唐天华[1] 任海全[1] 张之勇[1] 赵海涛[1] 范华[1] 乔高娟[1] 姜国胜[1] 

机构地区：[1]山东省医学科学院基础医学研究所,山东省现代医用药物与技术重点实验室,山东济南250062

出　　处：《中国实验血液学杂志》2008年第5期1030-1034,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金,编号30771103;山东省卫生厅青年基金2007年计划项目(2007QZ023);科技部中瑞政府间合作项目,编号AM15B1;山东省优秀中青年科学家基金项目,编号03BS033;山东省自然科学基金重点项目,编号Z2004C08;山东省卫生高层次人才1020工程专项经费资助(鲁卫科教国合字2008-2);山东省科技攻关重点项目(2007GG2002023);山东省自然科学基金项目(Y2005C39)

摘　　要：本研究探讨六亚甲基二乙酰胺(HMBA)对髓系白血病HL-60细胞分化的作用及其分子机制。应用MTT比色法观察不同浓度HMBA在不同时间点对细胞增殖的抑制作用,采用Wright-Giemsa染色观察细胞形态学变化,运用流式细胞仪测定细胞分化抗原CD11b的表达,并进行细胞周期分析,半定量RT-PCR分析c-myc、mad1、p21、p27、hTERT、HDAC1基因mRNA的表达。结果表明:HL-60细胞经不同浓度(0.5、1、2mmol/L)HMBA处理后细胞增殖受到较明显的抑制,并随时间延长、剂量增大抑制作用明显增强。HL-60细胞经2mmol/L HMBA处理后,细胞形态学上趋于分化成熟,细胞停滞于G0/G1期;CD11b表达显著增高;c-myc、hTERT mRNA表达显著下调,mad1、p21、p27mRNA表达显著上调,而HDAC1mRNA表达无明显变化。结论:HMBA能诱导HL-60细胞分化,其分子机制可能是通过调节c-myc/mad1开关引起p21、p27上调,并且下调hTERT mRNA表达,与HDAC1活性抑制无关。The objective of this study was to investigate the effect of hexamethylene bisacetamid （HMBA） on differentiation of HL-60 cells and its possible molecular mechanism. HL-60 cells were co-cultured with different concentrations of HMBA （0.5, 1, 2 mmol/L） for 4 days, then the proliferation was assayed by MTT at different time points. Wright-Giemsa staining was used to observe the change in morphology. Cell differentiation antigen CDllb expression and the altered distribution of cell cycle in HL-60 induced by HMBA were analyzed by flow cytometry. The expressions of c-myc,madl, p21, p27, hTERT and HDAC1 mRNA were detected by RT-PCR. The results indicated that the proliferation of HL-60 cells was inhibited by HMBA in a time-and-dose-dependent manner. Upon 2 mmol/L HMBA treatment, the HL-60 cells arrested at G0/G1 phase and differentiated into granular line in morphology, with the up-regulation of CDllb expression. The expression of c-myc and hTERT mRNA obviously down-regulated, the expression of p21, p27 and madl mRNA up-regulated, while there was no change of the expression of hTERT mRNA. It is concluded that effect of HMBA on the differentiation of HL-60 cells may partly contribute to switch from c-myc to madl expression, to up-regulate expressions of p21 and p27 mRNA, and down-regulate hTERT mRNA expression, while there is no relation with the expression of HDACI mRNA.

关 键 词：六亚甲基二乙酰胺 HL-60细胞 细胞周期 细胞增殖 细胞分化 

分 类 号：R979.1[医药卫生—药品] R733.7[医药卫生—药学]