雄黄诱导急性早幼粒细胞白血病维甲酸耐药细胞株NB4-R1细胞凋亡的蛋白质组学研究  

Proteomic study of retinoid acid resistant NB4-R1 cells apoptosis induced by realgar

作　　者：齐珺[1] 张梅[1] 贺鹏程[1] 王鸿丽[2] 王新阳[3] 

机构地区：[1]西安交通大学医学院第一附属医院血液科,710061 [2]军事医学科学院国家生物医学分析中心 [3]西安交通大学医学院第一附属医院泌尿外科实验室

出　　处：《中华血液学杂志》2010年第11期752-757,共6页Chinese Journal of Hematology

摘　　要：目的 获得并鉴定雄黄（四硫化四砷,As4S4）诱导急性早幼粒细胞白血病（APL）维甲酸耐药细胞株NB4-R1细胞凋亡的差异蛋白质组.方法 运用高分辨二维双向电泳获得As4S4诱导NB4-R1细胞凋亡前后的蛋白质组表达谱,通过凝胶图像分析筛选出差异蛋白,经胶内酶解后应用MALDI-TOF-MS、MALDI-TOF/TOF、UPLC-MS/MS串联质谱技术和生物信息学鉴定差异蛋白.结果 共筛选鉴定出22个表达升高2倍以上或降低50%以上的蛋白质点,最终获得21种已知蛋白,包括在As4 S4作用后表达下调的5种蛋白,分别为SET/I2PP2A、RPP2、PCBP1、EIF4H-1和ANP32A/I1PP2A;在As4S4作用24 h后表达上调的2种蛋白,分别为HSP 27和HMGB1;及在As4S4作用48 h后表达上调的14种蛋白质,分别为PHB、ERP29、DNAJC8、PSMB4、ACTB、RPP0、RhoGDI2、α-tubulin、Transcription factor、RBM15/OTT1、eIF5A1和H2B1M等.结论 筛选鉴定出As4 S4诱导NB4-R1细胞凋亡过程中有21种蛋白差异表达.其中SET、RPP2和PHB可能成为治疗维甲酸耐药APL的新靶点.Objective To obtain and identify the differential proteome of apoptosis induced by realgar （tetra-arsenic tetra-sulfide, As4S4 ） in retinoid acid （RA） resistant human acute promyelocytic leukemia （APL） cell line NB4-R1 cells. Methods The comparative proteomic expressive profiles of NB4-R1 cells treated with and without As4S4 were obtained with the high-resolution two-dimensional electrophoresis system.After the analysis of ImageMasterTM 2D Platinum software combining artificial comparison. The differential expression proteins were screened and performed in-gel digestion and extraction of peptides, applied mass spectrometry MALDI-TOF-MS, MALDI-TOF/TOF, UPLC-MS/MS and bioinformatics to identify differential expressive proteins. Results Twenty-two spots with more than 2-fold （ ≥2 or ≤0.5 ） expression changes were identified and 21 known proteins obtained, including 5 down-regulated （ SET/I2PP2A, RPP2, PCBP1,EIF4H-1 and ANP32A/I1PP2A） after exposed to As4S4,2 up-regulated （ HSP27 and HMGB1 ） after exposed for 24 h, and 14 up-regulated （PHB, ERP29, DNAJC8, PSMB4, ACTB, RPP0, RhoGDI2, alpha-tubulin,transcription factor, RBM15/OTT1, eIF5A1 and H2B1M et al. ） after exposed for 48 h. Conclusion It is the first time to successfully obtain and identify the global proteome of apoptosis induced by As4S4 in R4A-resistant cells. Among them, expressional and functional regulation of target proteins SET, RPP2 and PHB might be the potential novel therapeutic target for RA-resistant APL.

关 键 词：白血病 早幼粒细胞 急性 耐药性 维甲酸 细胞系 NB4一R1 四硫化四砷 细 胞凋亡 蛋白质组 

分 类 号：R686[医药卫生—骨科学]