摘要
目的 探讨组蛋白去乙酰化酶抑制剂三丁酸甘油酯 (TB)体外诱导白血病细胞株SHI 1生长抑制、分化和凋亡作用 ,并对其作用机制进行初步研究。方法 应用细胞计数、锥虫蓝拒染法观察TB对细胞的生长抑制作用 ,通过细胞形态学观察、NBT还原实验、细胞表面抗原CD11b和CD14的检测、Annexin标记、DNA凝胶电泳等方法分析TB对SHI 1细胞的诱导分化及凋亡作用。通过West ernblot方法及逆转录聚合酶链反应检测TB作用前后细胞组蛋白H3乙酰化水平以及 p2 1WAF1/CIP1表达量的改变。结果 ①TB可以抑制SHI 1细胞的增殖及活力 ,呈剂量 时间依赖关系。②TB 0 .1mmol/L可诱导SHI 1细胞发生部分分化 ,NBT阳性率升高 ,细胞表面CD11b、CD14表达上调。③SHI 1经TB 0 .5～ 1.0mmol/L作用 4 8h ,出现典型的凋亡形态学改变。细胞AnnexinⅤ结合力明显上升 ,并可见典型的DNA梯形条带。TB作用后SHI 1细胞组蛋白H3乙酰化水平升高以及 p2 1WAF1/CIP1表达上调。结论 TB能抑制SHI 1细胞的增殖并影响细胞活力 ,诱导SHI 1细胞分化和凋亡 ,其机制与TB引起组蛋白乙酰化水平升高、继而上调 p2 1WAF1表达有关。
Objective To investigate the effects of tributyrin (TB), a histone deacetylase inhibitor, on the growth, differentiation and apoptosis of SHI 1 leukemia cells and explore its possible mechanism. Method Cell proliferation and viability were determined by cell counting, trypan blue dye exclusion. Cell morphological analysis, Annexin binding, DNA electrophoresis, expression of CD11b and CD14, NBT reduction were performed to evaluate differentiation and apoptosis of SHI 1 cells. The level of acetylated histone H3 was detected by Western blot and p21 WAF1/CIP1 expression by semi quantitative RT PCR. Results TB inhibited the proliferation and viability of SHI 1 cells in a time dose dependent manner. The morphology of SHI 1 cells cultured in the presence of 0.1 mmol/L TB for 72 hs was more mature with higher NBT positivity and up regulated expressions of CD11b and CD14 than that of control group. Exposed to 0.5-1.0 mmol/L TB for 48 hs, SHI 1 cells exhibited the morphological hallmarks of apoptosis, the increasing of Annexin binding and the DNA ladder on gel electrophoresis. The level of acetylated histone H3 and p21 WAF1 mRNA extracted from SHI 1 cells were increased by the treatment of TB. Conclusion TB can inhibit proliferation, induce differentiation and apoptosis of SHI 1 cells. The mechamism may associate with its up regulation of acetylated histone and the expression of p21 WAF1 .
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2004年第11期662-665,共4页
Chinese Journal of Hematology
基金
江苏省卫生厅资助项目 (H980 1)
