Stathmin和CrkL蛋白在耐阿霉素的K562白血病细胞中表达上调  

Up-regulation of Stathmin and CrkL Protein Expressions in Adriamycin-resistant Leukemia Cell Line K562/A02

作　　者：徐建萍[1,2] 胡建达[1] 林敏辉[1] 李静[1] 刘庭波[1] 

机构地区：[1]福建医科大学附属协和医院血液病研究所福建省血液病学重点实验室,福建福州350001 [2]福建医科大学医学技术与工程学院检验系,福建福州350004

出　　处：《中国实验血液学杂志》2011年第6期1383-1387,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金(编号30572132);福建省自然科学基金(编号C0410029);福建医科大学基金(编号FJGXY04022)

摘　　要：本研究应用蛋白质组学方法比较白血病K562细胞及其耐阿霉素的K562/A02细胞中蛋白质表达谱的差异,筛选新的耐药相关蛋白。利用荧光差异双向电泳(2D-DIGE)技术分离K562细胞及K562/A02细胞的总蛋白,用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/MS)对差异表达蛋白质点进行鉴定,搜索蛋白质数据库,获得差异蛋白质的信息,并用蛋白免疫印迹法和实时荧光定量PCR法在蛋白质水平和mRNA水平对差异蛋白质进行验证。结果显示,经过DIGE技术和质谱分析,鉴定出8个蛋白质在耐阿霉素的K562细胞中表达有差异,其中2个表达下调,6个表达上调,它们分别参与细胞能量代谢、细胞增殖、细胞凋亡、细胞信号转导和基因转录等。在表达上调的蛋白中挑选Stathmin和CrkL蛋白,在K562及其耐药细胞株中进行了蛋白免疫印迹实验,结果与双向电泳结果一致;实时荧光定量PCR显示,CrkL在mRNA水平变化与在蛋白质水平变化一致,而Stathmin在mRNA水平变化与蛋白质水平变化不完全一致。结论:白血病K562细胞及其耐阿霉素K562/A02细胞蛋白质表达谱有差异,Stathmin和CrkL蛋白可能与耐药有关,值得进一步探讨。The purpose of this study was to compare the differences of the protein expression profiles between human myeloid leukemia K562 cells and adriamycin-resistant K562/A02 cells,as well as to select novel resistance-related proteins in myeloid leukemia by means of proteomics.The total cellular proteins were separated from K562 and adriamycin-resistant K562/A02 cells by using technigue of two dimensional difference in gel electrophoresis（2D-DIGE）.Differentially expressed proteins were analyzed by matrix-assisted laser desorption ionization/time of flight-mass spectrometry（MALDI-TOF/MS）,and by protein database searching.Moreover,the differentially expressed proteins were verified at protein and mRNA levels by Western blot assay and quantitative real time PCR.The results showed that 8 proteins differentially expressed in adriamycin-resistant K562/A02 cells,among them 2 proteins were identified to be down-regulated and 6 to be up-regulated.These identified proteins involved in the cell energy metabolism,cell proliferation,cell apoptosis,signal transduction,gene transcription and translation respectively.The results assayed by Western blot were similar to those detected by 2D-PAGE.Two up-regulated proteins Stathmin and CrkL were selected for verification in K562 and K562/A02 cells.As a result,the results detected by Western blot were identical with results from 2D-DIGE;real time quantitative PCR assay showed that the changes of CrkL at mRNA level were identical with changes at protein level,but no complete identity of Stathmin changes at mRNA level and protein level was observed.It is concluded that the difference of protein expression profile exists in K562 and K562/A02 cells.Stathmin and CrkL proteins may be involved in the drug resistance and suggest a novel clue for the resistant mechanisms in myeloid leukemia,which is worth further to explore.

关 键 词：STATHMIN蛋白 CrkL蛋白 阿霉素 多药耐药 K562细胞 K562/A02细胞 

分 类 号：R738.1[医药卫生—肿瘤]