Han Dong-Mei, Chen Xie-Qun, Liang Rong, Dong Bao-Xia, Yin Wen
Department of Hematology, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2006 Feb;14(1):21-4.
To investigate the influence of sodium valproate (VPA) on proliferation, apoptosis and differentiation of HL-60 cell line, effect of VPA in various concentrations on proliferation of HL-60 cells was detected by MMT; Wright-Giemsa staining was performed to observe the morphologic changes of HL-60 cells; NBT experiment was used to test the differentiation of HL-60 cells; flow cytometry was used to observe cell cycles and analyze the apoptosis. The results indicated that the changes of the growth curve showed inhibition of proliferation of HL-60 cells. After a 24-48 hours culture with 2 mmol/L VPA, the cells exhibited nuclear shrinkage, pyknosis fragmentation and appearance of apoptosis bodies. The percentage of the annexin V(+)/PI(-) cells which were apoptotic increased from 2.9% to 17.1%; hypodiploid peak was observed; the percentage of HL-60 cells in G(1) phase increased from 51.1% to 84.6% and the cells in S phase decreased from 37.9% to 14.4%. After a week culture with 0.25 mmol/L VPA, the cells exhibited characteristics of differentiation. The percentage of NBT positive cells was (47 +/- 2)%. It is concluded that VPA can inhibit the proliferation of HL-60 cells while inducing differentiation and apoptosis of these cells. The mechanism needs to be further studied.
为研究丙戊酸钠（VPA）对HL-60细胞系增殖、凋亡及分化的影响，采用MTT法检测不同浓度VPA对HL-60细胞增殖的影响；进行瑞氏-吉姆萨染色观察HL-60细胞的形态学变化；采用NBT实验检测HL-60细胞的分化情况；运用流式细胞术观察细胞周期并分析凋亡情况。结果表明，生长曲线变化显示HL-60细胞的增殖受到抑制。用2 mmol/L VPA培养24 - 48小时后，细胞出现核固缩、核碎裂及凋亡小体。凋亡的膜联蛋白V(+)/碘化丙啶(-)细胞百分比从2.9%增至17.1%；观察到亚二倍体峰；HL-60细胞G(1)期百分比从51.1%增至84.6%，S期细胞从37.9%降至14.4%。用0.25 mmol/L VPA培养一周后，细胞呈现分化特征。NBT阳性细胞百分比为(47±2)%。结论是VPA可抑制HL-60细胞增殖，同时诱导这些细胞分化和凋亡。其机制有待进一步研究。
