The localization of beta 1,4-galactosyltransferase and its significance for malignancy

β1,4-半乳糖基转移酶的定位及其对恶性肿瘤的意义

• 批准号: 04671018
• 负责人: AOKI Daisuke
• 金额: $ 0.45万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671018/
• 关键词: Galactosyltransferase; Tumor marker; Ovarian cancer; Immunofluorescence; Duble-determinant EIA; Immuno-dlectronmicroscopy; Site^directed mutagenesis; Double‐determinant EIA; double-determinant EIA

In this study, 1) usefulness of galactosyltranseferase associated with tumor (GAT) as a tumor marker for ovarian tumors, 2) subcellular localization of galactosyltransferase(GalT), and 3) the mechanism for thetention of Galt in Golgi apparatus were investigated. Their outlines were described as follows ; 1) In order to measure GAT with double-denterminant enzyme immunoassay, we utilized monoclonal antibodies(Mabs) terned Mab8513 and Mab8628 generated against the GalT extracted from ascites of ovarian cancer patients. Assay of the serum-GAT levels showed favorite rate (52.9%) in ovarian cancers and extremely low false positive rate (5.7%) in benign tumors, indicating GAT to ve useful in terms of very low specificity for ovarian cancers. 2) Histochemical analysis using Mab8628 demonstrated a strong positive reaction in the suprasuclear area of cacer cells which coincided with trans-side of the Golgi apparatus as observed with immunoelectron microscopy. 3) Immunohistochemical expmination using Cos-1 cells transfected with wild and mutated cDNA for human galacrosyltransferase demonstrated that amino acid residues, Cys and His in a membrane-anchoring domain are necessaty for GalT to reside on the Golgi.

本研究探讨了肿瘤相关半乳糖基转移酶（GAT）作为卵巢肿瘤标志物的作用，半乳糖基转移酶（GalT）的亚细胞定位，以及GalT在高尔基体中的滞留机制。1）为了用双抗体夹心酶联免疫法检测卵巢癌患者腹水中的GAT，我们制备了抗卵巢癌患者腹水中GAT的单克隆抗体Mab 8513和Mab 8628。血清GAT检测卵巢癌的阳性率为52.9%，良性肿瘤的假阳性率为5.7%，提示GAT对卵巢癌的诊断特异性较低。2)用Mab 8628进行的组织化学分析表明，在cacer细胞的核上区有强阳性反应，这与免疫电镜观察到的高尔基体的反侧相吻合。3)用转染人半乳糖基转移酶野生型和突变型cDNA的Cos-1细胞进行的免疫组化实验表明，膜锚定结构域中的氨基酸残基Cys和His是GalT驻留在高尔基体上所必需的。

项目成果

D.Aoki et al: "The role of tumor marker in the cancer screning : with special reference to uterine endometrial and ovarian cancer." Cancer Res. Clinic. 1. 3-14 (1992)

D.Aoki 等人：“肿瘤标志物在癌症筛查中的作用：特别是子宫内膜癌和卵巢癌。”

野澤志朗: "卵巣癌における癌関連ガラクトース転移酵素(GAT)の基礎的・臨床的検討(第一報)" 癌と化学療法. 20. 271-277 (1993)

Shiro Nozawa：“卵巢癌中癌症相关半乳糖基转移酶 (GAT) 的基础和临床研究（首次报告）”《癌症与化疗》20. 271-277 (1993)。

Nozawa,S.: "CA54/61 as a marker for epithelial ovarian cancer." Cancer Research. 52. 1205-1209 (1992)

Nozawa,S.：“CA54/61 作为上皮性卵巢癌的标志物。”

野澤志朗: "卵巣癌における癌関連ガラクトース転移酵素(GAT)の基礎的・臨床的検討(第二報)" 癌と化学療法. 20. 279-285 (1993)

Shiro Nozawa：“卵巢癌中癌症相关半乳糖基转移酶 (GAT) 的基础和临床研究（第二份报告）”《癌症与化疗》20. 279-285 (1993)。

青木 大輔: "がんのスクリーニングにおける腫瘍マーカーの意義 -子宮体癌および卵巣癌を中心に-" Cancer Research Clinic(CRC). 1(4). 3-14 (1992)

Daisuke Aoki：“肿瘤标志物在癌症筛查中的意义 - 关注子宫内膜癌和卵巢癌 -”癌症研究诊所 (CRC) 1(4) (1992)。