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[维甲酸耐药与敏感急性早幼粒细胞白血病细胞间差异蛋白质表达谱分析]

[Analysis of differential protein expression profile between retinoic acid resistant and sensitive acute promyelocytic leukemia cells].

作者信息

Qin Hui, Liu Ting, Yang Jin-Liang, Huang Xin, Liu Bin, Song Xin, Zhao Xia, Wei Yu-Quan

机构信息

Department of Hematology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China.

出版信息

Ai Zheng. 2006 Jul;25(7):828-32.

Abstract

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.

METHODS

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.

RESULTS

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.

CONCLUSION

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.

摘要

背景与目的

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。

方法

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。

结果

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。

结论

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。

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