Identification of Tumor-associated Autoantigens for the Diagnosis of Colorectal Cancer in Serum Using High Density Protein Microarrays
Identification of Tumor-associated Autoantigens for the Diagnosis of Colorectal Cancer in Serum Using High Density Protein Microarrays
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DOI：
10.1074/mcp.m800596-mcp200
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发表时间：
2009-10-01
影响因子：
7
通讯作者：
Ignacio Casal, J.
中科院分区：
文献类型：
--
作者：
Babel, Ingrid;Barderas, Rodrigo;Ignacio Casal, J.
有越来越多的证据表明存在与癌症进展相关的自身抗体。抗体是血清筛选的首选靶标，因为其稳定性和灵敏免疫测定的适用性。通过使用含有8000种人类蛋白质的商业蛋白质微阵列，我们检测了20例结直肠癌（CRC）患者和健康受试者的血清，以确定自身抗体模式和相关抗原。在蛋白质微阵列中，43种蛋白质被肿瘤血清和参考血清差异识别（p值< 0.04）。五种免疫反应性抗原，PIM 1，MAPKAPK 3，STK 4，SRC和FGFR 4，在癌症样品中显示出最高的流行率，而ACVR 2B在正常血清中更丰富。其中三个，PIM 1，MAPKAPK 3和ACVR 2B，用于进一步验证。通过免疫印迹和组织芯片免疫组化证实了这些抗原在CRC细胞系和结肠粘膜上的表达水平显著增加。基于MAPKAPK 3和ACVR 2B蛋白的组合的诊断ELISA在使用包含代表不同进展阶段和对照受试者的94份血清的独立样品组后，对于CRC区分分别产生73.9%和83.3%的特异性和灵敏度值（曲线下面积，0.85）。总之，这些研究证实了CRC特异性自身抗体的存在，并揭示了新的疾病个体标志物（PIM 1，MAPKAPK 3和ACVR 2B），其诊断CRC的特异性和灵敏度高于先前报道的血清生物标志物。Molecular & Cellular Proteomics 8：2382-2395，2009.
There is a mounting evidence of the existence of autoantibodies associated to cancer progression. Antibodies are the target of choice for serum screening because of their stability and suitability for sensitive immunoassays. By using commercial protein microarrays containing 8000 human proteins, we examined 20 sera from colorectal cancer (CRC) patients and healthy subjects to identify autoantibody patterns and associated antigens. Forty-three proteins were differentially recognized by tumoral and reference sera (p value < 0.04) in the protein microarrays. Five immunoreactive antigens, PIM1, MAPKAPK3, STK4, SRC, and FGFR4, showed the highest prevalence in cancer samples, whereas ACVR2B was more abundant in normal sera. Three of them, PIM1, MAPKAPK3, and ACVR2B, were used for further validation. A significant increase in the expression level of these antigens on CRC cell lines and colonic mucosa was confirmed by immunoblotting and immunohistochemistry on tissue microarrays. A diagnostic ELISA based on the combination of MAPKAPK3 and ACVR2B proteins yielded specificity and sensitivity values of 73.9 and 83.3% (area under the curve, 0.85), respectively, for CRC discrimination after using an independent sample set containing 94 sera representative of different stages of progression and control subjects. In summary, these studies confirmed the presence of specific autoantibodies for CRC and revealed new individual markers of disease (PIM1, MAPKAPK3, and ACVR2B) with the potential to diagnose CRC with higher specificity and sensitivity than previously reported serum biomarkers. Molecular & Cellular Proteomics 8: 2382-2395, 2009.