论文部分内容阅读
为筛选肺腺癌(lung adenocarcinoma,AdC)发病相关蛋白质,首先采用激光捕获显微切割技术(laser capture microdissection,LCM)分别从AdC组织和正常支气管上皮(normal bronchial epithelium,NBE)组织中切割并收集AdC细胞和NBE细胞,再应用双向凝胶电泳技术(two-dimensional gel electrophoresis,2-DE)分离经LCM收集的细胞蛋白质,通过PDQuest软件分析差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异表达蛋白质,组织芯片免疫组化方法检测差异蛋白质膜联蛋白A4(annexin A4)在30例AdC组织、配对的癌旁组织和淋巴结转移癌组织中的表达水平。研究结果显示,通过蛋白质组学方法建立了LCM收集的AdC和NBE细胞的2-DE图谱,质谱鉴定得到了33个差异表达蛋白质,其中21个蛋白质在AdC细胞中表达上调,12个蛋白质在AdC细胞中表达下调。组织芯片免疫组化结果显示,与癌旁肺组织相比,annexin A4在AdC组织中的表达水平显著上调,且在AdC淋巴结转移癌组织中的表达明显高于其原发癌组织。上述结果提示annexin A4与AdC的发病及淋巴结转移相关,有望成为诊断AdC及预测AdC转移的分子标志物。
To screen for the pathogenesis-related proteins of lung adenocarcinoma (AdC), they were first excised from AdC tissue and normal bronchial epithelium (NBE) tissue by laser capture microdissection (LCM) AdC cells and NBE cells, and the cell proteins collected by LCM were separated by two-dimensional gel electrophoresis (2-DE). Differentially expressed protein spots were analyzed by PDQuest software. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry The differentially expressed proteins were identified by MALDI-TOF-MS, and the protein expression of annexin A4 was detected by immunohistochemistry in 30 AdC tissues, matched paracancerous tissues and lymph node metastasis tissues. The results showed that 2-DE maps of AdC and NBE cells collected by LCM were established by proteomics. 33 differentially expressed proteins were identified by mass spectrometry. 21 of them were up-regulated in AdC cells and 12 in AdC Down-regulation of cells. Tissue microarray immunohistochemistry results showed that annexin A4 expression was significantly up-regulated in AdC tissues compared with adjacent lung tissues, and was significantly higher in AdC lymph node metastases than in primary cancerous tissues. These results suggest that annexin A4 is associated with the pathogenesis of AdC and lymph node metastasis, which is expected to become a molecular marker for the diagnosis of AdC and the prediction of AdC metastasis.