融合细胞株Eahy926与亲本人肺腺癌细胞株A549差异表达蛋白的蛋白质组学分析

融合细胞株Eahy926是人肺腺癌细胞株A549和人脐静脉内皮细胞杂交而成的永生化细胞株，具有血管内皮细胞的特性，已广泛用于内皮细胞相关研究. 本研究应用蛋白质组学技术分析融合细胞株Eahy926与亲本人肺腺癌细胞株A549的蛋白质差异表达，探讨融合细胞生物学特性变化及其机制. 对Eahy926和亲本A549的细胞总蛋白质进行双向凝胶电泳，在PDQuest软件辅助下找出差异表达蛋白质点，经肽质量指纹图谱(peptide mass fingerprinting, PMF) 和串级质谱(tandem mass spectrometry,TMS) 分析，SWISS-PROT数据库检索，成功鉴定出28个差异蛋白，如CATB、CK8、CK18、annexin A2、GRP78、HSP90、HSP60、vimentin等一些与分子伴侣、氧化应激、能量代谢、信号转导等有关，并与肿瘤细胞分裂增殖、分化凋亡、侵袭转移、免疫逃逸以及肿瘤血管生长密切关联的蛋白质. 研究发现,融合细胞株Eahy926和人肺腺癌细胞株A549的蛋白质组表达谱存在明显差异，这将有助于今后进一步探讨肿瘤细胞与内皮细胞的相互作用机制及其融合细胞的特性，筛选肿瘤增殖和转移相关蛋白质及分子标志物，亦可为肿瘤的抗血管生成治疗提供新思路.

Proteomic Analysis of Differential Expression Proteins in Fusion Cell Line Eahy926 and Parent Lung Adenocarcinoma Cell Line A549

The human endothelial-like Eahy926 cell line,derived from fusion of human umbilical vein endothelial cells with lung adenocarcinoma cell line A549,has the characteristics of vascular endothelial cells.To investigate the biological difference between Eahy926 and the parental A549 cells,we compared the overall proteome between these two cell lines.Total protein lysates of the two cell lines were extracted and separated by 2-DE,and the image analysis was performed using PDQuest.After searching against SWISS-PROT protein database,total of 28 differentially expressed proteins was successfully identified by peptide mass fingerprinting(PMF)and tandem mass spectrometry(TMS).Fifteen proteins were found to be overexpressed in Eahy926 cells,while thirteen proteins were found to be overexpressed in A549 cells.These molecules belong to diverse intracellular functional pathways such as molecular chaperone activity,oxidative stress,energy metabolism and signal transduction,and some are particularly involved in proliferation,differentiation,apoptosis,invasion and metastasis of cancer cells.The data in this study show signifcant differential proteomic expression pattern exists between A549 cell line and Eahy926 cell line,providing interesting evidence to understand the interaction between cancer cells and endothelial cells,the characteristics of vascular endothelial cells,and the mechanisms of lung cancer invasion and metastasis.