Qin Hui, Liu Ting, Yang Jin-Liang, Huang Xin, Liu Bin, Song Xin, Zhao Xia, Wei Yu-Quan
Department of Hematology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China.
Ai Zheng. 2006 Jul;25(7):828-32.
BACKGROUND & OBJECTIVE: This study was to compare the protein expression profiles between retinoic acid (RA) resistant and sensitive acute promyelocytic leukemia (APL) cells by proteomic research method.
Total cellular proteins extracted from a RA sensitive cell line NB4 and a RA resistant cell line MR2 were separated by two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE). High quality 2D-PAGE protein profiles were obtained and analyzed by PDQuest v7.1 analysis software to screen differentially expressed protein spots. Those sports were identified by mass spectrometry.
2-DE patterns of APL cell lines with high-resolution and reproducibility were obtained. The average spots for MR2 and NB4 cells were 890+/-45 and 912+/-56, respectively. 57 significantly differentially expressed protein spots were screened, among which 23 protein spots were identified to be up-regulated and 34 down-regulated in MR2 cells compared with NB4 cells. Ten proteins were identified by mass spectrometry, with a successful identification rate of 70%. The identified proteins could be classified into different categories: oncogenes,cell cycle regulator and signal transducer.
The utilization of 2D-PAGE is effective in identifying the protein expression profiles between RA resistant and sensitive APL cells, therefore this study may provide a novel clue to elucidate the drug resistant mechanisms of all-trans retinoic acid.
本研究旨在通过蛋白质组学研究方法比较维甲酸（RA）耐药和敏感的急性早幼粒细胞白血病（APL）细胞之间的蛋白质表达谱。
从RA敏感细胞系NB4和RA耐药细胞系MR2中提取的总细胞蛋白，通过二维（2D）聚丙烯酰胺凝胶电泳（PAGE）进行分离。获得高质量的2D-PAGE蛋白质谱，并使用PDQuest v7.1分析软件进行分析，以筛选差异表达的蛋白质斑点。这些斑点通过质谱进行鉴定。
获得了具有高分辨率和可重复性的APL细胞系的2-DE图谱。MR2和NB4细胞的平均斑点数分别为890±45和912±56。筛选出57个显著差异表达的蛋白质斑点，其中与NB4细胞相比，MR2细胞中有23个蛋白质斑点上调，34个下调。通过质谱鉴定了10种蛋白质，成功鉴定率为70%。鉴定出的蛋白质可分为不同类别：癌基因、细胞周期调节因子和信号转导子。
二维聚丙烯酰胺凝胶电泳（2D-PAGE）的应用有效地鉴定了RA耐药和敏感的APL细胞之间的蛋白质表达谱，因此本研究可能为阐明全反式维甲酸的耐药机制提供新线索。
