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Development of serological diagnosis of pancreatic cancer for mass screening

用于大规模筛查的胰腺癌血清学诊断的发展

基本信息

批准号：
63870029
负责人：
KAWAI Keiichi
金额：
$ 3.39万
依托单位：
Kyoto Prefectural University of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63870029/
关键词：
Pancreatic cancer gamma-GTP Mass screening Monoclonal antibody 血清診断 α-GTP γーGTP

项目摘要

Two different types of gamma-glutamyltranspeptidase (gamma-GTP) have been found in normal human pancreas following bromelain treatment. On the other hand, three human pancreatic ductal cell carcinomas have only a single type of gamma-GTP upon analysis with polyacrylamide gel electrophoresis, anion-exchange column chromatography and isoelectric focusing. Carcinoma gamma-GTPs were almost identical to one of the two types of normal pancreatic gamma-GTPs. The gamma-GTP from pancreatic carcinomas bound to anion-exchange column and was eluted at the same Nacl fractions as normal pancreatic gamma-GTP. The properties of pancreatic carcinoma gamma-GTP, as assessed by binding to concanavalin A and lentil lectin affinity columns, were also similar to one of the two enzymes of normal pancreas. However, E-PHA lectin column chromatography demonstrated differences in their elution pattern. Compared with normal pancreatic gamma-GTPs, the elution pattern of the carcinoma gamma-GTP was slowly delayed by the E-PHA lectin column. This finding suggests the presence of a bisecting N-acetyl-glucosamine-rich sugar chain in pancreatic carcinoma gamma-GTP as well as the possible presence of a different sugar structure in the carcinoma gamma-GTP. Then we immunized Balb/c mice with those gamma-GTPs and the spleen cells from immunized mice fused with myeloma cell line. The hybridoma cells were screened for novel gamma-GTP by ELISA method and also the monoclonal antibodies were examined for specificity by the method of immunohistochemistry using human pancreatic cancer tissues. In future we are planning to use these developmental monoclonal antibodies for the mass screening of pancreatic cancer.

菠萝蛋白酶处理后，在正常人胰腺中发现了两种不同类型的γ-谷氨酰转肽酶（γ-GTP）。另一方面，三个人胰腺导管细胞癌只有一个单一类型的γ-GTP的分析与聚丙烯酰胺凝胶电泳，阴离子交换柱层析和等电聚焦。癌γ-GTP与两种正常胰腺γ-GTP之一几乎相同。胰腺癌的γ-GTP与阴离子交换柱结合，并在与正常胰腺γ-GTP相同的NaCl组分下洗脱。胰腺癌γ-GTP的性质，通过结合刀豆球蛋白A和小扁豆凝集素亲和柱进行评估，也类似于正常胰腺的两种酶之一。然而，E-PHA凝集素柱层析证明了它们的洗脱模式的差异。与正常胰腺γ-GTP相比，癌γ-GTP的洗脱模式被E-PHA凝集素柱缓慢延迟。这一发现表明胰腺癌γ-GTP中存在一条富含N-乙酰基-葡萄糖胺的二等分糖链，并且癌γ-GTP中可能存在不同的糖结构。然后用这些γ-GTP免疫Balb/c小鼠，并将免疫小鼠的脾细胞与骨髓瘤细胞系融合。用ELISA方法筛选新型γ-GTP的杂交瘤细胞，并用人胰腺癌组织免疫组化方法检测单克隆抗体的特异性。在未来，我们计划使用这些发展单克隆抗体的大规模筛查胰腺癌。