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《工程(英文)》 >> 2021年 第7卷 第10期 doi: 10.1016/j.eng.2021.04.026

从结直肠癌细胞和组织样本释放的外泌体中检测肿瘤相关抗原自身抗体的多路生物传感诊断平台展示了对结直肠癌的高诊断能力

a Chronic Disease Program (UFIEC), Carlos III Health Institute, Madrid E-28220, Spain
b Department of Biochemistry and Molecular Biology, Faculty of Optics and Optometry, Complutense University of Madrid, Madrid E-28040, Spain
c Gastroenterology Unit, San Carlos Clinical Hospital, Madrid E-28040, Spain
d Surgical Pathology Department, San Carlos Clinical Hospital, Madrid E-28040, Spain
e Translational Oncology Division, OncoHealth Institute, Jimenez Diaz Foundation University Hospital, Madrid E-28040, Spain
f Department of Biochemistry, Faculty of Medicine, Alberto Sols Institute of Biomedical Research, CSIC-UAM, Madrid E-28029, Spain
g Department of Analytical Chemistry, Faculty of Chemical Sciences, Complutense University of Madrid, Madrid E-28040, Spain
h Center for Biological Research, CSIC, E-28040 Madrid, Spain
i Molecular Pathology and Therapeutic Targets Group, La Paz University Hospital (IdiPAZ), Madrid E-28046, Spain

收稿日期: 2020-09-30 修回日期: 2021-02-05 录用日期: 2021-04-07 发布日期: 2021-08-14

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摘要

结直肠癌(CRC)是全球导致癌症相关死亡的第二大原因。CRC患者的5年生存率与诊断分期息息相关。CRC早期诊断患者的5年生存率高于80%,CRC晚期诊断患者的5年生存率低于10%。大量研究表明,患者血清中特定CRC自身抗原[肿瘤相关抗原(TAA)]对应的自身抗体有助于早期诊断。因此,本文旨在识别CRC特异性自身抗体的自身抗原靶点,借助液体活检技术有效筛查CRC患者和健康个体。为此,我们从CRC患者体内提取5个CRC细胞系和组织样本,通过免疫沉淀-质谱分析法测量其分泌的外泌体的蛋白质含量,共鉴定出103种蛋白质为潜在的CRC特异性自身抗原。采用生物信息学技术和荟萃分析,我们选定了15种与实际CRC自身抗原类似度更高的蛋白质,以便后续借助蛋白质印迹法(WB)和免疫组织化学技术(IHC)评估它们在CRC预后中的作用。结果发现,在患者的组织和血浆外泌体样本中,有11种蛋白质发生蛋白质水平失调,有9种蛋白质与CRC预后有关。经验证发现,除一例外,所有研究均显示出具有统计学意义的高诊断能力,采用荧光Halotag磁珠,或者借助多路生物传感平台(磁性微载体为固相载体,由用于CRC细胞检测的共价固定Halotag融合蛋白修饰),能够有效筛查区分CRC患者、癌前病变个体与健康个体。综上所述,本文的研究结果突出了此研究方法在识别慢性疾病特征TAA时的有效性;此外,本文采用的测量血浆中TAA对应的自身抗体水平的方法可以应用到具有高诊断能力的CRC检测即时医疗(POC)设备中。

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