异硫氰酸苯己酯诱导伊马替尼耐药K562/G01细胞株凋亡的组蛋白调控机制研究  

Study on the Effects and Histone Modulative Mechanisms of Apoptosis by PHI in Imatinib-resistant K562/G01 Cell Line

作　　者：吴荣娟[1] 黄轶群[1] 马旭东[1] 

机构地区：[1]福建医科大学附属漳州市医院血液科,漳州363000

出　　处：《福建医科大学学报》2012年第6期380-384,共5页Journal of Fujian Medical University

基　　金：卫生部科学研究基金-福建省卫生教育联合攻关计划项目(WKJ2008-2-55);福建医科大学科学研究发展专项基金计划项目(FZS08018);福建省漳州市2010科技计划项目(Z2010080)

摘　　要：目的体外观察异硫氰酸苯己酯(PHI)对人慢性粒细胞白血病伊马替尼耐药的K562/G01细胞株的凋亡的影响,初步探讨其可能的组蛋白调控机制。方法流式细胞仪检测PHI作用前后K562/G01细胞凋亡的变化;Western Blot检测PHI处理K562/G01细胞株后凋亡相关蛋白Bcl-2,procaspase-3,组蛋白H3、H4乙酰化状态,组蛋白H3K4、H3K9甲基化状态的变化。结果 PHI可以诱导K562/G01细胞凋亡,且呈浓度依赖性。PHI终浓度为0,10,20,40μmol/L时,K562/G01细胞凋亡率分别为(3.76±1.46)%,(8.89±2.31)%,(18.10±3.56)%,(35.35±3.70)%,差别有统计学意义(n=3,P<0.05);凋亡相关蛋白Bcl-2及procaspase-3的表达下降;组蛋白H3、H4乙酰化及组蛋白H3K4甲基化水平均上升,而H3K9甲基化水平下降,随着作用时间的延长变化趋势更加明显。结论 PHI能诱导K562/G01细胞凋亡,可能与PHI对组蛋白的乙酰化及甲基化调控有关。Objective To study the effects on apoptosis by PHI in K562/G01 cell line and to ex- plore its potential histone modulative mechanisms. Methods AnnexinV/PI assays were used to evaluate cell apoptosis in K562/G01 cell lines. The expression levels of Bcl-2 and procaspase-3 proteins,the chan- ges of histone acetylated H3 and H4, histone methylated H3K4 and H3K9 in K562/G01 cells treated with PHI were detected by Western Blot. Results PHI induced apoptosis in K562/G01 cells in a concentra- tion-dependent manner. Along with the concentration adding, the ratio of apoptosis was raising. The control group, 10 μmol/L PHI group, 20 μmol/L PHI group and 40 μmol/L PHI group, the correspond- ingapoptosis ratio was （3.76±1.46）%, （8. 89±2. 31）%, （18. 10±3. 56）%, （35. 35±3. 70）%, the difference was statistically significant （P〈0.05）. PHI down-regulated proteins of Bcl-2, procaspase 3 and induced an accumulation of histone acetylated H3, H4, and increased histone methylation H3K4 and decreased methylated H3K9. Conclusions PHI could induce apoptosis in K562/G01 cell line. The effects may be associated with the modulations of histone acetylation and methylation.

关 键 词：白血病 髓系 慢性 BCR—ABL阳性 硫 氰化物 苯 细胞系 肿瘤 甲基化 乙酰化作用 细胞凋亡 组蛋白类 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]