丙戊酸诱导白血病U937细胞凋亡机理的实验研究  

Empirical Study on the Mechanism of the Apoptosis Induced by VPA

作　　者：薛红漫[1] 郭海霞[1] 李晓圆[1] 张金华[1] 李文益[1] 

机构地区：[1]中山大学附属第二医院儿科,510120

出　　处：《实用癌症杂志》2006年第5期458-460,共3页The Practical Journal of Cancer

摘　　要：目的从分子水平探索丙戊酸(VPA)诱导白血病U937细胞凋亡的机制,为临床治疗提供理论依据。方法将U937细胞分为1mmol/LVPA作用组、1mmol/LVPA+1μmol/LzVAD-fmk作用组和空白对照组,作用72h后进行Annixin-ⅴ及PI双重染色,在流式细胞仪上检测细胞凋亡;采用流式细胞仪检测处理前后各组细胞Bcl-2、Bax和Bcl-xl的平均荧光指数(MFI)以及胱冬肽酶(caspase)3、8和9的含量。结果1mmol/LVPA诱导U937凋亡率为(75.78±4.20)%(P<0.01);多cas-pase抑制剂zVAD-fmk可全部抑制U937凋亡,凋亡率为(2.89±0.36)%(P<0.01)。细胞内Bcl-2、Bax和Bcl-xl的MFI无明显变化。VPA作用后U937中caspase3由(14.09±1.19)%上升至(32.30±2.47)%,caspase8由(4.58±1.41)%上升至(86.47±3.26)%(P均<0.01),caspase9变化不显著。结论VPA通过激活caspase3和caspase8诱导U937凋亡;VPA诱导U937凋亡并非通过改变细胞中Bcl-2等的含量实现。Objective The aim of this survey is to investigate the mechanism of VPA inducing the apotosis of leukemic cells on the molecular level. Methods The leukemic cell lines U937 were divided into three groups （control group; 1 mmol/L VPA group and 1 mmol/L VPA ＋ 1 μmol/L zVAD-fmk group）. 72 hours after treated, cells were double stained with Annixin-V and PI, then was analyzed with the FCM to detect apoptosis. Before and after treated the mean fluorescent index （MFI） of Bcl-2、Bax、Bcl-xl and the levels of caspase 3、8、9 in cells were also detected with the FCM. Results 1mmol/L VPA induced the apoptosis of U937, the apoptotic rate of U937 was （75.78±4.20）%. Compared with the control group there were significant difference （P 〈 0.01）; Pancaspase inhibitor zVAD-fmk could fully inhibit the apoptosis of U937, and the apoptotic rate was （2.89±0.36）% （P 〈 0.01）. The MFI of Bcl-2、Bax and Bcl-xl in the cell line had not changed significantly （P 〉 0.05）. After treated with VPA, the level of caspase 3 in U937 was from （14.09±1.19） % up to （32.30±2.47） %, and caspase 8 was from （4.58±1.41） % up to （86.47±3.26） % （both P 〈 0.01）, but there was no significant change in caspase 9. Conclusion VPA induce the apoptosis of U937 through the activation of caspase 3 and 8; and it does these without the alteration of the expression of Bcl-2、Bax and Bcl-xl.

关 键 词：丙戊酸 白血病 BCL-2蛋白 胱冬肽酶 

分 类 号：R733.7[医药卫生—肿瘤]