LRRC4基因转染U251细胞的比较蛋白质组学分析  

Comparative Proteomics Analysis of Differentially Expressed Proteins in U251 Cells Transfected by pcDNA3.1(+) or LRRC4 Gene

作　　者：杨静[1] 武明花[1] 王蓉[1] 张祖萍[1] 李小玲[1] 李桂源[1] 

机构地区：[1]中南大学肿瘤研究所,长沙410078

出　　处：《中国生物化学与分子生物学报》2009年第4期345-351,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展规划(973计划;No.2006CB910504;2006CB910502);国家自然科学基金(No.30770825);111计划(No.111-2-12);湖南省自然科学基金(No.06JJ20080)~~

摘　　要：LRRC4是我室自主克隆的一个脑组织优势表达基因.前期研究结果表明,外源性LRRC4基因转染至U251细胞,可明显地抑制U251细胞的增殖、黏附、趋化和侵袭等生物学行为.因此,LRRC4亦是一个脑胶质瘤抑制性基因.为了进一步了解LRRC4在胶质瘤发生发展中的调控作用,本研究采用荧光差异凝胶电泳(2D-DIGE)和质谱分析技术获得了LRRC4转染U251细胞的11个差异表达蛋白质,并用Western印迹证实了U251细胞在转染LRRC4基因前后热休克蛋白27、stathmin1和S100钙结合蛋白A11的差异表达变化.这些差异蛋白质涉及细胞代谢、增殖、转录、信号转导等众多事件,表明LRRC4基因转染U251细胞后可能通过调控这些蛋白质的表达而参与细胞的增殖、黏附、趋化和侵袭等生物学过程.LRRC4 is a dominantly expressed gene involved in neural development and axon growth in the brain, and its expression is also associated with glioma progression. As the glioma clinical grading increases, the expression of LRRC4 gradually decreases, or even diminished. LRRC4 transfection can inhibit the proliferation, adhesion, chemotactie response and invasion in U251. cells, therefore, referred as a tumor suppressive gene in glioma. In this study, we used differential in-gel electrophoresis （DIGE） and mass spectrometry （MS） to analyze the proteins in U251 cells transfected by pcDNA3.1 （ ＋ ） or LRRC4, and discovered 11 differentially expressed proteins, including heat shock protein 27, stathmin and S100 calcium binding protein All that were validated by Western blot. These differentially expressed proteins were involved in diversified biological activities, such as cell metabolism, proliferation, translation and signal transduction, etc. The results indicated that the involvement of LRRC4 in the onset and progression of glioma nfight be related to its regulation of those differentially expressed proteins.

关 键 词：LRRC4 胶质瘤 荧光差异凝胶电泳 MALDI-TOF/TOF-MS/MS 

分 类 号：R73-3[医药卫生—肿瘤] R739[医药卫生—临床医学]