蛋白质组学方法分析PRL-3功能相关蛋白  

Comparative proteomics analysis of PRL-3 functionally related protein

作　　者：关玉峰[1] 刘璐[1] 伍衡[1] 来伟[1] 李守峰[1] 褚忠华[1] 

机构地区：[1]中山大学孙逸仙纪念医院胃肠外科,广州510288

出　　处：《中华实验外科杂志》2011年第6期944-946,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：广东省社会发展计划基金资助项目（20078031513012）;广东省自然科学基金资助项目（9151008901000058、10151008901000071）

摘　　要：目的蛋白质组学方法分析转染肝再生磷酸酶-3（PRL-3）后结肠癌细胞kVo的蛋白表达改变。方法构建绿色荧光蛋白载体PAcGFP—C3-PRL-3并转染至结肠癌细胞LoVo中，获得稳定表达GFP—PRL-3的结肠癌细胞LoVo—PRL-3。Westernblot检测转染后细胞的PRL-3表达。采用双向凝胶电泳（2-DE）分离LoVo—PRL-3细胞及转染空载体PAcGFP—C3的LoVo-Control细胞总蛋白，从中选取差异表达蛋白质点，通过基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF-TOF-MS）鉴定差异表达的蛋白质。结果Westernblot结果显示PRL-3在LoVo—PRL-3细胞中高表达，而未检测到在LoVo—Control细胞中表达。通过双向凝胶电泳图谱分析发现20个差异蛋白质斑点，质谱成功鉴定出17种蛋白。与LoVo—Control细胞比较，10种蛋白在LoVo-PRL-3细胞中高表达，而7种蛋白在 LoVo—PRL-3细胞中表达降低。结论成功鉴定出PRL-3功能相关蛋白，为进一步研究PRL-3促进结直肠癌转移机制奠定基础。Objective To identify the protein expression patterns after the colon cancer cells LoVo transfected with phosphatase of regenerating liver-3 （ PRL-3 ） using a comparative proteomic approach. Methods The green fluorescent protein vector PAcGFP-C3-PRL-3 was constructed and transfected into the colon cancer cells LoVo and colon cancer cells stably expressing GFP-PRL-3 （ LoVo-PRL-3 ） were established. Western blotting was used to detect the expression levels of PRL-3 in LoVo-PRL-3 cells. 2-DE was applied to separate the whole proteins in LoVo-PRL-3 cells and LoVo control cells that trasfected with vectors PAcGFP-C3. The proteins with statistically significant differential expression between LoVo-PRL-3 and LoVo control ceils were identified by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry （ MALD I-TOF-TOF-MS）. Results Western blotting showed that PRL-3 was highly ex- presssed in LoVo-PRL-3 cells, but undetectable in LoVo control cells. Twenty proteins spots were found as differentially expressed proteins using two-dimensional gel electrophoresis and 17 proteins were successfully identified using mass spectrometry. Among these identified proteins, 10 proteins were significantly up-regu- lated, while 7 proteins were notably down-regulated in the lovo-PRL-3 compared to the lovo control cells. Conclusion The PRL-3 functionally related proteins were successfully identified, which laying the foundation for further studies on PRL-3-promoted metastasis.

关 键 词：结肠癌 蛋白质组学 肝再生磷酸酶-3 

分 类 号：R735.35[医药卫生—肿瘤]