Functional analysis of carbohydrate antigens in chemically induced tumor cells derived from beta-1,4-galactosyltransferase-I knockout mice

β-1,4-半乳糖基转移酶-I 敲除小鼠化学诱导肿瘤细胞中碳水化合物抗原的功能分析

• 批准号: 15500298
• 负责人: HASHIMOTO Noriyoshi
• 金额: $ 2.37万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15500298/
• 关键词: Carbohydrate; Knockout mice; Galactosyltransferase; Tumor antigen; Galactose; Chemical carcinogen; Skin tumor; Malignancy

Beta-1,4-galactosyltransferase I (beta4GalT-I) is an essential glycosyltransferase to synthesize some kinds of type 2 N-glycans and core 2 O-glycans. We have generated beta4GalT-I knockout (KO) mice to study the multiple in vivo function of these carbohydrates. First, we have generated embryonic fibroblast cell lines from both beta4GalT-I homozygously mutated and heterozygously mutated, phenotypically normal, mice. Both cell lines were immortalized by repetitive passages. Transfection of mouse beta4GalT-I expression vector to beta4GalT-I null fibroblast cell line have no effect on immortalization of cells. In order to evaluate tumor specific carbohydrate antigens such as sialyl Lewis antigens in tumorigenesis, we have conducted functional analysis of carbohydrate antigens using chemically induced skin tumor cell lines derived from beta4GalT-I KO mice. No significant differences in cellular growth and adherent abilities to fibronectin were observed between beta4GalT-I null cells and those which were transfected with mouse beta4GalT-I expression vector. When motility and invasiveness of beta4GalT-I null cell lines through fibronectin-coated and/or Matrigel-coated transwells were assessed, significant migrations and invasions were observed. Moreover, the cell motility and invasiveness were declined by the transfection of mouse beta4GalT-I expression vector in an expression level dependent manner. Though sialyl Lewis antigens have not be detected after beta4GalT-I transfections to beta4GalT-I null cell lines, significant expressions of galactose residuedetected by RCA 120 lectin in the beta1,4-linkage was observed. These results suggest that carbohydrates synthesized by beta4GalT-I regulate the malignancy of tumor cells.

β-1，4-半乳糖基转移酶I(beta4GalT-I)是合成多种N-糖链和核糖链所必需的糖基转移酶。我们已经建立了Beta4GalT-I基因敲除(KO)小鼠来研究这些碳水化合物在体内的多重功能。首先，我们已经从表型正常的Beta4GalT-I纯合子突变和杂合突变的小鼠中建立了胚胎成纤维细胞系。两种细胞系都通过重复传代而永生化。将小鼠Beta4GalT-I表达载体导入Beta4GalT-I缺失的成纤维细胞系，对细胞永生化无影响。为了评价肿瘤特异性碳水化合物抗原如唾液酸路易斯抗原在肿瘤发生中的作用，我们用化学诱导的Beta4GalT-I KO小鼠皮肤肿瘤细胞系对碳水化合物抗原进行了功能分析。未转染鼠β4GalT-I表达载体的细胞生长和对纤维连接蛋白的黏附能力与转染鼠β4GalT-I表达载体的细胞相比无明显差异。当评估通过纤维连接蛋白包被和/或Matrigel包被的跨孔的Beta4GalT-I缺失细胞系的运动性和侵袭性时，观察到显著的迁移和侵袭。此外，小鼠Beta4GalT-I表达载体以表达水平依赖的方式降低细胞的运动能力和侵袭力。虽然将β4GalT-I导入Beta4GalT-I缺失细胞系后未检测到唾液酸化Lewis抗原，但用RCA120凝集素检测到β-1，4-链上有明显的半乳糖残留量。这些结果表明，由Beta4GalT-I合成的碳水化合物调节肿瘤细胞的恶性。

项目成果

Analysis of human IgA nephropathy-like disease in galactosyltransferase KO mice.

半乳糖基转移酶 KO 小鼠中人 IgA 肾病样疾病的分析。

• 发表时间: 2004
• 期刊: Nephrology (Carlton) 9 Suppl 2
• 作者: Nishie T;Miyaishi O;Naruse C;Hashimoto N;Asano M.
• 通讯作者: Asano M.

Characterization of serum IgA in beta4GalT-I-deficient mice developing IgAN-like disease

发生 IgAN 样疾病的 beta4GalT-I 缺陷小鼠血清 IgA 的特征

• 发表时间: 2005
• 期刊: Nephrology Suppl 6
• 作者: Asano;M. et al.
• 通讯作者: M. et al.

Impaired selectin ligand biosynthesis and reduced inflammatory responses in β-1,4-galactosyltransferase-I-deficient mice.

β-1,4-半乳糖基转移酶-I 缺陷小鼠的选择素配体生物合成受损并减少炎症反应。

• 发表时间: 2003
• 期刊: Blood 102
• 作者: Asano;M. et al.
• 通讯作者: M. et al.

Impaired selectin-ligand biosynthesis and reduced inflammatory responses in beta-1,4-galactosyltransferase-I-deficient mice.

β-1,4-半乳糖基转移酶-I 缺陷小鼠的选择素配体生物合成受损并减少炎症反应。

• 发表时间: 2003
• 期刊: Blood 102(5)
• 作者: Asano M;Nakae S;Kotani N;Shirafuji N;Nambu A;Hashimoto N;Kawashima H;Hirose M;Miyasaka M;Takasaki S;Iwakura Y.
• 通讯作者: Iwakura Y.

Characterization of serum IgA in beta4GalT I-deficient mice developing IgAN-like disease.

发生 IgAN 样疾病的 beta4GalT I 缺陷小鼠血清 IgA 的特征。

• 发表时间: 2005
• 期刊: Nephrology (Carlton) 10 Suppl 6
• 作者: Asano M;Nishie T;Miyaishi O;Azuma H;Kameyama A;Naruse C;Hashimoto N;Yokoyama H;Narimatsu H;Wada T.
• 通讯作者: Wada T.