摘要
目的:探讨丙戊酸钠(VPA)通过抑制组蛋白脱乙酰化酶(HDAC)的活性,消除AML1-ETO融合蛋白转录抑制作用,抑制Kasumi-1细胞增殖及诱导凋亡的机制。方法:不同浓度VPA处理Kasumi-1细胞不同时间后,应用台盼蓝拒染人工计数法观察VPA对细胞增殖的影响,普通光学显微镜观察细胞形态改变,流式细胞术分析细胞周期和检测髓系分化抗原CD11b的表达水平的变化,DNA凝胶电泳检测细胞的凋亡。结果:VPA能显著抑制Kasumi-1细胞的增殖,且呈剂量和时间依赖性,VPA处理Kasumi-1细胞72h时的半数抑制浓度(IC50)为2.33mmol/L;VPA处理后,细胞周期检测G0/G1期细胞比例逐渐增高;VPA能诱导髓系分化抗原CD11b表达水平的阳性率升高,呈时间及剂量依赖性;VPA能诱导Kasumi-1细胞凋亡,Kasumi-l细胞经VPA3mmol/L处理72h后,出现典型的凋亡细胞所具有的梯形DNA条带。结论:VPA能抑制Kasumi-1细胞增殖,阻滞细胞周期于G0/G1期,诱导细胞部分分化和凋亡。
Objective： To study the mechanism of histone deacetylase inhibitor VPA repressing HDAC activity, reversing the transcription inhibition effect of AML1-ETO fusion protein, inhibiting cell growth and inducing apoptosis of Kasumi- 1 cell. Methods： Trypan blue refusal staining method was used to detect the proliferation of Kasumi-1 cell treated with VPA, common light microscope was applied to observe the morphology change of the cell, flow cytometry was done to analyze cell cycle and detect the expression level of myeloid cell differentiation antigen, and DNA gel electrophoresis was used to observe cell apoptosis. Results： The proliferation of Kasumi-1 cell was obviously inhibited by VPA on dose- and time-dependent manner. The IC50 was 2.33 mmol/L treated with VPA for 72 h. Incubation with VPA,the cell ratio at G0/G1 phase increased. VPA induced myeloid cell differentiation antigen CD1 lb to increase with a time-and dose-dependent way and caused apoptosis. Wright＇ s staining and light microscope were used to show the morphology of partial differentiation and obvious apoptosis with 3 mmol/L VPA for 72 hours. The typical DNA ladder of apoptosis characteristic was observed with DNA gel electrophoresis. Gonclusion：VPA treatment can inhibit cell proliferation of Kasumi-1,and induce partial differentiation, apoptosis and accumulation of cells in G0/G1 phase.
出处
《天津医药》
CAS
北大核心
2008年第6期410-413,共4页
Tianjin Medical Journal
基金
河北省科技发展项目(项目编号:05276114)
