【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。更多还原

【Abstract】 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 CD11b expression and the altered distribution of cell cycle in HL-60 induced by HMBA were analyzed by flow cytometry.The expressions of c-myc,mad1,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 CD11b expression.The expression of c-myc and hTERT mRNA obviously down-regulated,the expression of p21,p27 and mad1 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 mad1 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 HDAC1 mRNA.更多还原