目的 利用蛋白质组学技术建立人肺腺癌细胞系A549与正常支气管上皮细胞系HBE的差异蛋白质表达谱,并对部分在A549细胞中表达明显上调的蛋白质点进行鉴定.方法 提取A549和HBE细胞的总蛋白,进行双向凝胶电泳(2-DE),经图像分析识别差异表达蛋白质点,再应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定出部份在A549细胞中表达明显上调的蛋白质.结果 (1)分析两种细胞系的2-DE图谱,发现差异表达蛋白质点共256个,仅在A549细胞中表达的蛋白质点15个,仅在HBE细胞中表达的蛋白质点21个;(2)通过肽质量指纹图谱分析鉴定出在A549细胞中表达明显上调的7种蛋白质,分别是表皮生长因子受体(EGFR)、鼠双微粒体2蛋白(MDM2)蛋白、CD44蛋白、细胞骨架蛋白细胞角蛋白8(CK8)、不均一性核糖体蛋白H(hnRNP H)、热休克蛋白60(HSP60)、磷酸甘油酸变位酶1(PGAM1).结论 运用蛋白质组学技术成功获得了A549细胞与HBE细胞的2-DE图谱,并鉴定出部分在A549细胞中表达明显上调的蛋白质,为进一步筛选用于人肺腺癌诊断、治疗及预后评估的分子标志物奠定了一定的基础.%Objective To establish differential protein expression profile of human lung adenocarcinoma cell line A549 and normal human bronchial epithelial cell line HBE using proteomics technology, and to identify some of the protein spots whose expression were up-regulated obviously in A549 cells. Methods Total protein of A549 and HBE cells were extracted and subjected to two-dimensional polyacrylamide gel electrophoresis(2-DE). The differentially expressed protein spots were recognized by image analysis, and then the matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS) was employed to identify proteins whose expression were up-regulated obviously in A549 cells. Results (1) Comparatively analyzing the 2-DE protein patterns of the two types of cell lines, a total of 256 differentially expressed protein spots were found, with 15 spots differen tially expressed only in A549 cells and 21 spots only in HBE cells. (2) 7 kinds of protein including epidermal growth factor receptor (EGFR) , murine double minute 2 (MDM2) , CD44 , cytokeratin 8(CK8) , heterogeneous nuclear ribonucleoprotein H (hnRNPs H) , heat shock protein 60 (HSP60) and phosphoglycerate mutase 1(PGAM1) , were identified by peptide mass fingerprintin,and their expression were obviously up-regulated in A549 cells. Conclusion 2-DE protein patterns of A54 9 and HBE cells are successfully obtained by proteomic technology, and some of the proteins whose expression were up-regulated obviously in A549 cells are identified. These lay a foundation for further screening of molecular diagnostic, treatment and prognostic markers of lung cancer.
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