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目的:探讨组蛋白去乙酰化酶抑制剂丙戊酸钠(sodium valproate,VPA)体外诱导白血病细胞kasumi-1的分化和凋亡作用。方法:MTT实验和台盼蓝拒染法检测VPA对细胞的增殖抑制,流式细胞仪检测细胞分化和凋亡及进行细胞周期分析,蛋白质印迹方法检测组蛋白H3(lys9)的乙酰化水平变化及p21Waf1/Cip1蛋白的表达。结果:VPA可以抑制kasumi-1细胞增殖。VPA作用后,细胞周期阻滞在G0/G1期,该期细胞由(53.07±6.05)%增加到(78.97±5.82)%,P<0.05。在较低浓度(0.5mmol/L),VPA可以诱导kasumi-1细胞髓系分化,其表面抗原CD13表达从(20.47±0.21)%上升到(68.67±1.15)%,P=0.000。联合G-CSF后作用更为显著〔(87.80±1.21)%〕,P=0.000。在较高浓度(2mmol/L)下,VPA则诱导kasu-mi-1细胞凋亡,流式检测AnnexinⅤ结合力上升。VPA能够上调kasumi-1细胞组蛋白H3乙酰化水平,增加p21Waf1/Cip1蛋白的表达。结论:VPA能通过抑制HDAC活性,抑制细胞增殖,诱导kasumi-1细胞分化和凋亡。
AIM: To investigate the differentiation and apoptosis of kasumi-1 induced by sodium valproate (VPA), a histone deacetylase inhibitor, in vitro. Methods MTT assay and trypan blue exclusion method were used to detect the proliferation inhibition of VPA. Flow cytometry was used to detect the cell differentiation and apoptosis. The changes of acetylation level of histone H3 (lys9) were detected by Western blotting And p21Waf1 / Cip1 protein expression. Results: VPA can inhibit the proliferation of kasumi-1 cells. After VPA treatment, the cell cycle arrest was at G0 / G1 phase and the cell number increased from (53.07 ± 6.05)% to (78.97 ± 5.82)%, P <0.05. At a lower concentration (0.5 mmol / L), VPA induced the myeloid differentiation of kasumi-1 cells, and the surface antigen CD13 expression increased from (20.47 ± 0.21)% to (68.67 ± 1.15)%, P = 0.000. Combined effect of G-CSF was more significant 〔(87.80 ± 1.21)%〕, P = 0.000. VPA induced the apoptosis of kasu-mi-1 cells at higher concentration (2mmol / L), and the binding capacity of Annexin V increased by flow cytometry. VPA can upregulate acetylation of histone H3 in kasumi-1 cells and increase p21Waf1 / Cip1 protein expression. Conclusion: VPA can induce kasumi-1 cell differentiation and apoptosis by inhibiting HDAC activity, inhibiting cell proliferation.