丙戊酸对胶质瘤细胞株T98-G体外作用实验研究

Anti-tumor activity of 2-propylpentanoic acid on human glioma cell line

目的研究抗癫痫常用药物丙戊酸（2-propylpentanoic acid，VPA）临床治疗浓度对人脑胶质瘤细胞株T98-G增殖抑制、细胞周期及组蛋白乙酰化的影响，并探讨其意义。方法四甲基偶氮唑蓝（MTT）比色法检测VPA对胶质瘤细胞株T98-G的细胞毒性作用；流式细胞术检测其对细胞周期动力学及其对细胞凋亡的影响；Western blot法检测胶质瘤细胞株在VPA处理后乙酰化组氨酸H3（Acetyl—Histone H3）、乙酰化组氨酸H4（Acetyl—Histone H4）的表达量变化。结果丙戊酸对胶质瘤细胞株T98-G具有抑制增殖作用，随着药物浓度的增加，抑制作用增强；临床常用浓度（1．0mmol／L）VPA能够对细胞周期动力学产生影响，S期细胞减少，而G1期、G2／M期细胞增加；Aceyl—HistoneH3、Aceyl—His／oneH4蛋白表达量明显上调。结论VPA能够抑制胶质瘤细胞生长，引起细胞周期阻滞，其作用可能与其抑制组蛋白去乙酰化酶活性有关。

Objective To investigate the anti-tumor activity of 2-propylpentanoic acid（VPA）, a common anti-epilepsy drug at clinical safety dosage, and the effect on the expression of Acetyl-Histone H3 and H4 expression. Methods Human glioma cell line T98-G was treated by VPA at concentration of 0. 2, 0. 6,1.0,2. 0,2.7,3.6,4. 5 mmol/L for 1,2,3,5 and 7 days, respectively. The Cellular proliferation was determined by the methyl thiazolyl tetrazolimn（MTT） assay, and cell cycle arrest and cell apoptosis were observed by flow eytometry. The expression of Aeetyl-Histone Ha and H4 were detected by Western Blotting analysis. Results Obvious changes in cell proliferation was found in the glioma cell line T08-G afler treatment with VPA in respective concentration for 24 h, and in a dose- and time- dependent manner. G1 or G2/M phase arrest was induced by the clinical safety （dosage） concentration 1.0 mmol/L at 24 hours. Cells in S phase were decreased about 10% while cells in C-1 , G2/M phase were increased accordingly. The expression of Aeeyl-Histone H3 and H4 were significantly increased after treatment with 1.0 mmol/L VPA at 24 hours,and in a time-dependent manner. Conclusion VPA may inhibit growth of glioma cell line T98-G, induce both G1 and G2 phase arrest, which could be resulting from inhibition of histone deaeetylase activity.