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分析来鉴定结直肠癌患者的肿瘤特异性抗原 确定具有抗肿瘤抗体的各种类型胃肠道恶性肿瘤患者的数量 从结直肠癌细胞系 DLD-1 中分离出蛋白质。通过二维聚丙烯酰胺凝胶电泳和蛋白质印迹到 P 聚偏二氟乙烯膜上，与来自结直肠癌患者的血清进行免疫反应。进行凝胶内胰蛋白酶消化、肽提取和基质辅助激光解吸/电离飞行时间质谱分析并从胃肠道恶性肿瘤和其他结直肠疾病患者的血清中鉴定免疫原性蛋白质。对健康受试者进行了针对已识别蛋白质的反应性测试，其中一种免疫原性蛋白质被鉴定为 β-微管蛋白，在 16/54 的患者中检测到抗体。 (30%) 结直肠癌，6/16 (38%) 胃癌，2/10 (20%) 食管癌和 4/10 (40%) 胰腺癌，而 2/3 (67%) 结直肠腺瘤，0/2 结肠癌克罗恩病和 0/1 阑尾类癌只有一名健康受试者具有抗体，对于结肠直肠癌患者，特异性为 96%。抗β-微管蛋白抗体的存在与分化良好的病变相关（p<0.05），而与疾病阶段或血管和淋巴管侵袭的存在没有相关性。通过基于蛋白质组学的方法，我们成功地鉴定了β-微管蛋白。作为结直肠癌和其他胃肠道恶性肿瘤中的肿瘤特异性抗原，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志物。

项目成果

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

基于蛋白质组学的胃癌肿瘤相关抗原及其相应自身抗体的鉴定。

• DOI: 10.3892/or_00000719
• 发表时间: 2010-04-01
• 期刊: Oncology reports
• 影响因子: 4.2
• 作者: S. Tsunemi;T. Nakanishi;Y. Fujita;G. Bouras;Yoshiharu Miyamoto;A. Miyamoto;E. Nomura;T. Takubo;N. Tanigawa
• 通讯作者: N. Tanigawa

Proteomics-based identification of autoantibody against CDC25B as a novel serum marker in esophageal squamous cell carcinoma.

基于蛋白质组学的 CDC25B 自身抗体鉴定作为食管鳞状细胞癌的新型血清标志物。

• DOI: 10.1016/j.bbrc.2008.08.039
• 发表时间: 2008-10-24
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: Wan;Ge Zhang;Jun;J. Cao;Xian;Li;Man‐zhi Li;Libing Song;Wenlin Huang;M. Zeng
• 通讯作者: M. Zeng

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

基于蛋白质组学的胃癌肿瘤相关抗原及其相应自身抗体的鉴定

• 发表时间: 2024-09-14
• 作者: 常深 聡一郎

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
• 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
• 作者: Akihoto Ogata; Akira Wada; Yoshihisa Fujita; et al.
• 通讯作者: et al.