双水相纯化人肺腺癌和癌旁正常肺组织质膜蛋白质组差异分析  

Comparative Analysis of Plasma Membrane Proteome Lung Adenocarcinoma and Paraneoplastic Tissues Purified from Two-phase Partition

作　　者：梁旭俊 郭爱云[1] 陈主初[1] 卢光琇[2,3] 张鹏飞[1,2,3] 

机构地区：[1]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙410008 [2]中南大学生殖与干细胞工程研究所,长沙410078 [3]人类干细胞国家工程研究中心,长沙410205

出　　处：《中国生物化学与分子生物学报》2015年第2期182-190,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.81372516);国家重大新药创制(No.2012ZX09303013-006);湖南省发改委科技基本建设项目投资计划(No.2013-1199)资助项目~~

摘　　要：肿瘤标志物对于肺腺癌病人的临床诊断和预后具有重要意义.本研究根据临床诊断选取人肺腺癌组织和癌旁正常肺组织为研究对象,采用差速离心联合双水相法纯化组织细胞质膜,运用同位素标记相对和绝对定量技术结合高效液相色谱-串联质谱技术,鉴定出肺腺癌组织和癌旁正常肺组织的差异蛋白质41种.同癌旁正常肺组织相比,18个蛋白质在肺腺癌组织中表达上调,23个蛋白质在肺腺癌组织中表达下调.生物信息学分析发现,差异质膜蛋白质FLOT1、CAV1和ITGB1均处于蛋白质相互作用网络的重要位置,可能参与了肺腺癌相关信号转导.利用蛋白质印迹和免疫组织化学染色验证,差异蛋白质FLOT1、CAV1和ITGB1在肺腺癌织和癌旁正常肺组织的表达情况,其验证结果与蛋白质组学研究结果一致.研究结果对肺腺癌诊断标志物和肺腺癌癌变分子机理研究具有重要意义.Tumor markers are of great significance in the clinical diagnosis and prognosis for lung adenocarcinoma patients. In this study,lung adenocarcinoma tissues and paired paraneoplastic normal lung tissues（ PNLTs） were collected for investigation. Differential centrifugation and aqueous two-phase partition technology was used to purify the plasma membrane（ PM）. HPLC-MS / MS was used in combination with i TRAQ（ isobaric tag for relative and absolute quantitation） for analysis. Differentially expressed PM proteins of 41 in total between primary lung adenocarcinomas and PNLTs were identified,including 18 up-regulated and 23 down-regulated in cancer tissues. Bioinformatic analysis showed that the identified FLOT1,CAV1 and ITGB1 were involved in protein-protein interaction network in the signal transduction of lung adenocarcinoma. The expressions of FLOT1,CAV1 and ITGB1 were detected by Western blotting and immunohistochemistry for validation.

关 键 词：人肺腺癌 ITRAQ 质膜 定量蛋白质组学 

分 类 号：Q51[生物学—生物化学] Q71