非小细胞肺癌患者尿中Exosomes蛋白质组的差异表达分析

Analysis of Differentially Expressed Proteome in Urinary Exosome from Non-small Cell lung Cancer Patients

除血浆之外,尿液是另一种寻找潜在生物标志物的重要生物材料。本研究以200000×g超速离心法分离正常人和非小细胞肺癌(NSCLC)患者尿液中的Exosomes,运用1DSDS-PAGE对Exosomes蛋白质组进行分组,从电泳胶上切取正常组和疾病组的20～31kDa条带,胰蛋白酶酶解后,进行HPLC-CHIP-MS/MS分析,并通过UniProtKB/SWISS-PORT数据库搜索鉴定了24种蛋白质,其中在NSCLC患者尿液Exosomes蛋白质组中发现了8种差异表达蛋白,包括免疫球蛋白κ的3个片段、2种Ras相关蛋白、谷胱甘肽S转移酶A2、血清淀粉样P成分前体和磷脂酰乙醇胺结合蛋白1。

Urine provides an alternative to blood plasma as a potential source of disease biomarkers. Exosomes was separated by ultracentrifuge at 200000 g in normal persons and non-small cell lung cancer （ NSCLC） patients′ urine. For proteomic analysis of urinary exosome,1D sodium dodecylsulfonate-polyacrylate gel electrophoresis（ SDS-PAGE） was carries out and cut the gel 31 kDa-20 kDa bands in normal group and disease group′s. These gel blocks were subjected to in-gel trypsinization,and the extracted peptides were analyzed HPLC-CHIP-MS/MS. Approximately 24 unique proteins were identified in the UniProtKB/SWISSPORT. The difference expression proteins were found in urinary exosome from NSCLC patients,including three fragment of the immunoglobulin kappa,two kinds of Ras related proteins,glutathione S-transferase A2, serum amyloid P-component precursor and phosphatidylethanolamine-binding protein 1.