基于Label-free技术的非小细胞肺癌蛋白质差异研究

Protein Difference in Non-small Cell Lung Cancer Based on Label-free Technique

目的 基于非小细胞肺癌(non-small-cell lung cancer,NSCLC)及与其配对的非癌性邻近组织(non-cancerous adjacent tissues,NATs)的蛋白质组学分析,寻找NSCLC诊断相关蛋白标志物。方法 应用非标记定量蛋白组学(LFP)技术,以NSCLC患者配对的NATs为对照,对5例NSCLC患者进行癌组织蛋白质组学分析。以差异倍数>1.5(上调)或<0.67(下调)且P<0.05为标准筛选差异蛋白。应用String网站和R语言对差异蛋白进行基因本体论(gene ontology,GO)分析、京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析,为进一步研究提供良好基础。结果 本研究发现差异蛋白共644个,其中339个蛋白表达上调,305个蛋白表达下调。PCA结果显示,差异蛋白可明显区分NSCLC与NATs。GO分析结果表明,差异蛋白大多以细胞外泌体的形式存在且主要富集于调节胞外分泌、胞外分泌、骨髓细胞激活参与免疫反应等的生物学过程以及钙粘着蛋白绑定、细胞外基质绑定等的分子功能。KEGG分析结果显示,差异蛋白主要富集在抗坏血酸和醛酸代谢、组氨酸代谢、蛋白质消化吸收、丙酮酸代谢等通路(P<0.05)。结论 NSCLC与NATs存在明显差异,可为发现NSCLC新型标志物提供线索。

Objective Based on proteomic analysis of non-small-cell lung cancer(NSCLC) and non-cancerous adjacent tissues(NATs) with it,Search for protein markers associated with the diagnosis of NSCLC.Methods Unlabeled quantitative proteomics(LFP) technique was used to analyze the proteomics of cancer tissue in 5 patients with NSCLC,and the matched NATs of NSCLC patients were used as controls.The difference factor 1.5(up-regulated) or 0.67(down-regulated) and P<0.05 were used as the criteria to screen differential proteins.The gene ontology(GO) analysis and the Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis were conducted using the String web site and the R language.Genomes(Genomes) analysis It provides a good foundation for further study.Results A total of 644 different proteins were found in this study,among which 339 proteins were up-regulated and 305 proteins were down-regulated.PCA showed that differential proteins could significantly distinguish between NSCLC and NATs.The results of GO analysis showed that most of the differential proteins existed in the form of extracellular bodies and were mainly enriched in the biological processes of regulating extracellular secretion,extracellular secretion,activation of bone marrow cells and participation in immune response,as well as molecular functions of cadherin binding and extracellular matrix binding.KEGG analysis showed that differential proteins were mainly concentrated in ascorbic acid and uronic acid metabolism,histidine metabolism,protein digestion and absorption,pyruvate metabolism and other pathways(P<0.05).Conclusion There are significant differences between NSCLC and NATs,which can provide clues for the discovery of novel markers of NSCLC.