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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

基本信息

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

项目摘要

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.

通过对大肠癌细胞系蛋白质的质谱分析，鉴定大肠癌患者的肿瘤特异性抗原。确定显示抗肿瘤抗体的各种类型胃肠道恶性肿瘤患者的数量。用双向聚丙烯酰胺凝胶电泳分离大肠癌细胞系DLD-1的蛋白质，并将蛋白质印迹到P聚偏氟乙烯膜上，与大肠癌患者血清进行免疫反应。从考马斯亮蓝染色的凝胶中绘制并切除免疫反应性斑点。进行凝胶内胰蛋白酶消化、肽提取和基质辅助激光解吸/电离飞行时间质谱以鉴定免疫原性蛋白。检测来自胃肠道恶性肿瘤患者、其他结直肠疾病患者和健康受试者的血清对所鉴定蛋白质的反应性。其中一种免疫原性蛋白被鉴定为β-微管蛋白。结直肠癌、胃癌、食管癌和胰腺癌患者血清抗体阳性率分别为30%、38%、20%和40%，而结直肠腺瘤、克罗恩病和阑尾类癌患者血清抗体阳性率分别为67%、67%和67%。只有一名健康受试者有抗体，特异性为96%。对于那些患有结直肠癌的患者，抗β-微管蛋白抗体的存在与分化良好的病变相关（p<0.05），而与疾病分期或血管和淋巴管浸润的存在无关。通过基于蛋白质组学的方法，我们已经成功地鉴定了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和可测量的标志物。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

DOI：
发表时间：
2010
期刊：
影响因子：
0
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。

DOI：
发表时间：
2011
期刊：
Bulletin of the Osaka medical college Vol 57, No.1
影响因子：
0
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