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Expression cloning of carbohydrate sufotransferase and its implication in cancer metastasis

碳水化合物磺基转移酶的表达克隆及其在癌症转移中的意义

基本信息

批准号：
07670642
负责人：
HIRAIWA Nozomu
金额：
$ 1.54万
依托单位：
Aichi Cancer Center
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670642/
关键词：
glycosyltransferase sulfated glycolipid expression cloning hepatoma HNK-1 パンニング

项目摘要

We have tried to isolate a cDNA clone encoding human sulfotransferase, which could synthesize sulfated LacCer. In this research, we used a mammalian expression system with monoclonal antibodies which were established against sulfated glycolipids in our laboratory, and found that de novo expression of transgected cells. However, we are still in the way to get a cDNA that represent a tructural gene encoding a human sulfotransferase.At the same time, we have investigated expression pattern of the GalCer sulfotransferase which was recently cloned, among human liver and hepatoma cell lines. In Northern blots, the gene was expressed in human hepatoma cell lines in relation to product of sulfoglycolipids in these cells. This result might suggest that the enzyme encoded by the gene could synthesize sulfated LacCer, and this issue should be resolved upon analysis of enxymatic function derived from this gene.Among acidic glycolipids and glycoproteins, sialylated glyco-structures have function of cell adhesion like sulfated glycolipids, and we have investigated that expression of glycosyltransferases related to these products were actually regulatted in cancer-related and cell-specific condition. Especially some transferase is under the influence of the transcription activator of the oncovirus in synthesis of sialyl Le^x which mediates adhesion of leukocytes and leukemic cells to endothelium.

我们试图分离出一个编码人磺基转移酶的cDNA克隆，它可以合成硫酸化的LacCer。本研究利用本实验室建立的抗硫酸化糖脂单克隆抗体的哺乳动物表达系统，发现转染细胞的从头表达。同时，我们还研究了新近克隆的GalCer磺基转移酶在人肝细胞和肝癌细胞中的表达模式。在北方印迹中，该基因在人肝癌细胞系中表达，与这些细胞中的磺基糖脂产物有关。这一结果可能表明该基因编码的酶可以合成硫酸化的LacCer，这一问题应该通过分析该基因的酶学功能来解决。在酸性糖脂和糖蛋白中，唾液酸化的糖结构与硫酸化糖脂一样具有细胞粘附功能，并且我们已经研究了与这些产物相关的糖基转移酶的表达实际上在癌症相关的和细胞中受到调节，具体条件。特别是一些转移酶在肿瘤病毒的转录激活因子的影响下合成唾液酸Le^x，其介导白细胞和白血病细胞与内皮的粘附。