Li Yi-Qing, Yin Song-Mei, Feng Si-Qiong, Nie Da-Nian, Xie Shuang-Feng, Ma Li-Ping, Wang Xiu-Ju, Wu Yu-Dan
Department of Hematology, Sun Yat-Sen University, Guangzhou 510120, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Dec;18(6):1445-50.
This study was aimed to clarify whether valproic acid (VPA) induces apoptosis of leukemia HL-60 cell line and its possible mechanism. The effect of different concentrations and treatment time of VPA on HL-60 cell proliferation was assayed by cytotoxicity test (CCK-8 method) and fluorescence microscopy, and flow cytometry was used to detect cell apoptosis. The expressions of telomerase subunit h-tert mRNA and apoptosis-related protein as well as caspase-3 activity were detected by real time-quantitative PCR, Western blot and ELISA respectively. The results indicated that VPA inhibited proliferation of HL-60 cells and induced cell apoptosis in a dose dependent manner (r = -0.87). The expressions of anti-apoptotic protein BCL-2 and h-tert mRNA were significantly decreased while the pro-apoptotic protein BAX and caspase-3 activity increased after treatment with VPA. The apoptosis rate of HL-60 cell was negatively correlated with expression of h-tert mRNA. It is concluded that VPA can inhibit leukemia HL-60 cell proliferation and induce apoptosis. The VPA displays anti-leukemia activity possibly through reducing h-tert mRNA and BCL-2 protein expression, increasing BAX expression and activity of caspase-3.
本研究旨在阐明丙戊酸（VPA）是否诱导白血病HL-60细胞系凋亡及其可能机制。采用细胞毒性试验（CCK-8法）、荧光显微镜检测不同浓度及处理时间的VPA对HL-60细胞增殖的影响，并用流式细胞术检测细胞凋亡。分别采用实时定量PCR、Western印迹法及ELISA检测端粒酶亚基h-tert mRNA、凋亡相关蛋白的表达及caspase-3活性。结果表明，VPA抑制HL-60细胞增殖并呈剂量依赖性诱导细胞凋亡（r = -0.87）。VPA处理后，抗凋亡蛋白BCL-2和h-tert mRNA表达显著降低，而促凋亡蛋白BAX和caspase-3活性增加。HL-60细胞凋亡率与h-tert mRNA表达呈负相关。结论：VPA可抑制白血病HL-60细胞增殖并诱导凋亡。VPA可能通过降低h-tert mRNA和BCL-2蛋白表达、增加BAX表达及caspase-3活性发挥抗白血病活性。
