食管癌(EC)是一种致命的消化系统疾病,预后差,经常发生淋巴转移。然而,目前尚无用于EC诊断的可靠生物标志物.因此,我们使用无标记定量蛋白质组学图谱对来自8对EC患者的癌症和癌旁组织来源的外泌体进行了比较蛋白质组学分析,并通过生物信息学分析了差异表达的蛋白质.此外,使用纳米流式细胞术(NanoFCM)验证122例EC患者血浆来源外泌体的候选蛋白.在癌症和癌旁组织来源的外泌体中发现的803种差异表达蛋白中,686被上调,117被下调。细胞间粘附分子-1(CD54)被鉴定为上调的候选物,用于进一步研究,用免疫组织化学方法验证了其在EC患者癌组织中的高表达,实时定量PCR(RT-qPCR),和蛋白质印迹分析。此外,来自122例EC患者的血浆来源的外泌体NanoFCM数据与我们的蛋白质组学分析一致。接收机工作特性(ROC)分析表明,灵敏度,CD54的特异性值为0.702,66.13%,和71.31%,分别,用于EC诊断。小干扰(si)RNA用于沉默EC细胞中的CD54基因。一系列的化验,包括细胞计数试剂盒-8,粘附,伤口愈合,和Matrigel入侵,进行调查EC生存能力,粘合剂,迁徙,和侵入能力,分别。结果显示CD54促进EC增殖,迁移,和入侵。总的来说,组织来源的外泌体蛋白质组学有力地证明,CD54是EC诊断的有前景的生物标志物,也是EC发展的关键分子.
Esophageal cancer (EC) is a fatal digestive disease with a poor prognosis and frequent lymphatic metastases. Nevertheless, reliable biomarkers for EC diagnosis are currently unavailable. Accordingly, we have performed a comparative proteomics analysis on cancer and paracancer tissue-derived exosomes from eight pairs of EC patients using label-free quantification proteomics profiling and have analyzed the differentially expressed proteins through bioinformatics. Furthermore, nano-flow cytometry (NanoFCM) was used to validate the candidate proteins from plasma-derived exosomes in 122 EC patients. Of the 803 differentially expressed proteins discovered in cancer and paracancer tissue-derived exosomes, 686 were up-regulated and 117 were down-regulated. Intercellular adhesion molecule-1 (
CD54) was identified as an up-regulated candidate for further investigation, and its high expression in cancer tissues of EC patients was validated using immunohistochemistry, real-time quantitative PCR (RT-qPCR), and western blot analyses. In addition, plasma-derived exosome NanoFCM data from 122 EC patients concurred with our proteomic analysis. The receiver operating characteristic (ROC) analysis demonstrated that the AUC, sensitivity, and specificity values for
CD54 were 0.702, 66.13%, and 71.31%, respectively, for EC diagnosis. Small interference (si)RNA was employed to silence the
CD54 gene in EC cells. A series of assays, including cell counting kit-8, adhesion, wound healing, and Matrigel invasion, were performed to investigate EC viability, adhesive, migratory, and invasive abilities, respectively. The results showed that
CD54 promoted EC proliferation, migration, and invasion. Collectively, tissue-derived exosomal proteomics strongly demonstrates that
CD54 is a promising biomarker for EC diagnosis and a key molecule for EC development.