Identification of β-tubulin as a common immunogen in gastrointestinal malignancy by mass spectrometry of colorectal cancer proteome : implications for early disease detection

通过结直肠癌蛋白质组质谱分析鉴定β-微管蛋白作为胃肠道恶性肿瘤的常见免疫原：对早期疾病检测的影响

• 批准号: 21790545
• 负责人: FUJITA Yoshihisa
• 金额: $ 2.66万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21790545/
• 关键词: プロテオミクス; 癌診断マーカー; 自己抗体

To identify tumor-specific antigens in patients with colorectal cancer by mass spectrometry of cancer cell line proteins. To determine the number of patients with various types of gastrointestinal malignancy demonstrating anti-tumor antibodies. Proteins from the colorectal cancer cell line DLD-1 were separated by two-dimensional polyacrylamide gel electrophoresis and Western blotted onto P polyvinylidene fluoride membranes for immune reaction with sera from colorectal cancer patients. Immunoreactive spots were mapped and excised from Coomasie brilliant blue stained gels. In-gel trypsin digestion, peptide extraction and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was performed to identify immunogenic proteins. Sera from patients with gastrointestinal malignancies, other colorectal conditions and healthy subjects were tested for reactivity against identified proteins. One of the immunogenic proteins was identified as β-tubulin. Antibodies in patients were detected in 16/54 (30%) colorectal, 6/16 (38%) gastric, 2/10 (20%) esophageal and 4/10 (40%) pancreatic cancer, while in 2/3 (67%) colorectal adenoma, 0/2 colonic Crohn's and 0/1 carcinoid of the appendix. Only one healthy subject had antibodies making the specificity 96%. For those with colorectal cancer, the presence of anti-β-tubulin antibodies was associated with well differentiated lesions (p<0.05) while there were no correlations with disease stage or the presence of vascular and lymphatic invasion. Through a proteomics-based approach, we have successfully identified β-tubulin as a tumor-specific antigen in colorectal cancer and other gastrointestinal malignancies. Anti-β-tubulin antibodies may be a sensitive and measurable marker for early malignant disease.

通过癌细胞系蛋白的质谱法鉴定结直肠癌患者的肿瘤特异性抗原。确定患有抗肿瘤抗体的各种胃肠道恶性肿瘤的患者数量。通过二维聚丙烯酰胺凝胶电泳分离结直肠癌细胞系DLD-1的蛋白质，并将其蛋白质印迹到P聚偏二氟化物膜上，从结直肠癌患者的血清进行免疫反应。映射了免疫反应斑点，并从Coomasie亮蓝色染色凝胶中获得了极好的作用。进行凝胶胰蛋白酶消化，胡椒萃取和基质辅助激光解吸/电离飞行时间质谱法以鉴定免疫原性蛋白质。测试了胃肠道恶性肿瘤，其他有色疾病和健康受试者的血清针对已鉴定的蛋白质的反应性。一种免疫原性蛋白被确定为β-微管蛋白。在16/54（30％）结直肠癌中检测到患者的抗体，6/16（38％）胃，2/10（20％）食管和4/10（40％）胰腺癌，而2/3（67％）结直肠腺瘤，0/2 Colonial Crohn and Colonial Crohn's and 0/1/0/1 carcinix。只有一个健康的受试者具有使特异性为96％的抗体。对于那些结直肠癌的人，抗β-微管蛋白抗体的存在与良好的分化病变有关（P <0.05），而与疾病阶段或血管和淋巴浸润没有相关性。通过基于蛋白质组学的方法，我们成功地鉴定出β-微管蛋白是结直肠癌和其他胃肠道恶性肿瘤中肿瘤特异性抗原。抗β-微管蛋白抗体可能是早期恶性疾病的敏感且可测量的标记。

项目成果

Proteomics-based Identifying Autoantibody as a Novel Serum Marker in Esophageal Squamous Cell Carcinoma

基于蛋白质组学的识别自身抗体作为食管鳞状细胞癌的新型血清标志物

• 发表时间: 2010
• 作者: 田辺宏美;栗林景晶;辻直樹;渡邉直樹;藤田能久
• 通讯作者: 藤田能久

Proteomics-based identification of a tumor-associated antigen and its corresponding autoantibody in gastric cancer

• DOI: 10.3892/or_00000719
• 发表时间: 2010-04-01
• 期刊: ONCOLOGY REPORTS
• 影响因子: 4.2
• 作者: Tsunemi, Soichiro;Nakanishi, Toyofumi;Tanigawa, Nobuhiko
• 通讯作者: Tanigawa, Nobuhiko

ProLeomic analysis regarding resistance to anticancer drugs using a 5-fluorouracil-resitant human gastric cancer cell line and the radical-free and highly reducing method of two-dimensional electrophoresis.

使用5-氟尿嘧啶耐药人胃癌细胞系和无自由基、高度还原的二维电泳方法进行抗癌药物耐药性的ProLeomic分析。

• 发表时间: 2011
• 期刊: Bulletin of the Osaka medical college Vol 57, No.1
• 作者: AkihiLo Ogata;Akira Wads;Masami Ueta;Hideo FushiLani;SaLoru Tanaka;Kosei Kimura;Akiko Sakai;Hideji Yoshida;Akiko MiyamoLo;Yoshihisa Fujita;Nobuhiko Tanigawa
• 通讯作者: Nobuhiko Tanigawa

Proteomic analysis regarding resistance to anticancer drugs using a 5-fluorouracil-resitant human gastric cancer cell line and the radical-free and highly reducing method of two-dimensional electrophoresis

5-氟尿嘧啶耐药人胃癌细胞系抗癌药物耐药性的蛋白质组学分析及无自由基高还原二维电泳法

• 发表时间: 2011
• 期刊: Bulletin of the Osaka medical college
• 作者: Akihoto Ogata;Akira Wada;Yoshihisa Fujita;et al.
• 通讯作者: et al.