六亚甲基二乙酰胺对HL-60细胞细胞周期及其调节蛋白表达的影响  

Effects of Hexamethylene Bisacetamide on Cell Cycle and Expression of Its Regula tory Proteins in HL-60 Cells

作　　者：王钦红[1] 谢毅[1] 范华骅[2] 高砾[2] 刘燕[2] 

机构地区：[1]复旦大学附属华山医院血液科 [2]上海血液中心血液工程研究室,上海200051

出　　处：《中国实验血液学杂志》2003年第5期480-484,共5页Journal of Experimental Hematology

摘　　要：六亚甲基二乙酰胺 (hexamethylenebisacetamide ,HMBA)是一种极性诱导分化剂 ,已应用于临床治疗急性髓系白血病和骨髓增生异常综合征 (MDS) ,但其诱导髓系白血病细胞分化的机制尚不清楚。本研究观察了HMBA对HL 60白血病细胞细胞周期及其调节蛋白表达的影响 ,以探讨其药理机制。用流式细胞术检测HMBA对HL 60细胞细胞周期分布和细胞周期蛋白D、E及p2 7表达的影响 ,用RT PCR研究HMBA对细胞周期负性调控分子CKIp15,p16,p2 7基因mRNA表达的影响。结果表明 ,HMBA主要使HL 60细胞阻滞于G0 /G1期 ,经HMBA处理后HL 60细胞胞浆内细胞周期蛋白E蛋白表达显著降低 ,而细胞周期蛋白D和p2 7蛋白表达则显著增高 ;未经HMBA处理的HL 60细胞 p15,p16mRNA均无表达 ;3mmol/LHMBA处理 2 4小时后 p15mRNA出现弱阳性条带 ,48小时和 72小时后出现强阳性条带 ,p16mRNA则在 48小时后出现弱阳性条带 ,72小时后出现强阳性条带 ;p2 7mRNA在未经处理和处理后 2 4,48和 72小时后均出现强阳性条带 ,阳性程度无明显差异。结论 :HMBA的药理机制之一可能是通过下调细胞周期蛋白E ,上调p2 7蛋白的表达 ,同时使得 p15,p16基因mRNA重获表达 ,阻滞HL 60细胞周期于G1期 。Hexamethylene bisacetamide (HMBA) is referred as a differentiation-inducer for the clinical treatment of acute myeloid leukemia and myelodysplastic syndrome. H owever, the molecular mechanism of the effects of HMBA on myeloid leukemic cells remains unkown. In this study, the effects of HMBA on cell cycle and express ion of cell cycle regulatory proteins in HL-60 cell were investigated in order to explore its pharmacological mechanism. The altered distribution of cell cycle and expression of its regulatory proteins (cyclin D, cyclin E and p27) in HL-6 0 cell i nduced by HMBA were analyzed by flow cytometry. The effects on transcription for mRNA of CKI p15, p16 and p27 in HL-60 cell were further studied by RT-PCR. The results showed that HMBA could mainly commit HL-60 cell to G 0/G 1 arrest and the significantly decreased endocytic cyclin E protein and increased cyclin D/p27 protein after HMBA treatment were found. There was no expression of p15, p16 mRNA in untreated HL-60 cell and 3 mmol/L of HMBA could make them expressed after exposed for 24 h or 48 h respectively. The expression of p27 mRNA was positive and no obviously different in untreated HL -60 cell s exposed for 24 h, 48 h and 72 h. These results suggested that one of the phar macological mechanisms of HMBA was to elevate the expression of p27 and reduce t he cyclin E expression as well as to activate the expression of p15, p16 gene mRNA, that arrested cell at G 0/G 1 and exerted its effects of anti-prolifera tion.

关 键 词：六亚甲基乙酰胺 HL—60细胞 细胞周期 细胞周期调节蛋白 细胞周期蛋白 

分 类 号：R733.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]