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The effect of various in flammatory cytokines in Core2 glycosyltransferase

多种炎症细胞因子对Core2糖基转移酶的影响

基本信息

批准号：
09670587
负责人：
MAEMURA Kentarou
金额：
$ 1.09万
依托单位：
Second department of Internal medicine, Osaka Medical college
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670587/
关键词：
TGF-beta Core2 glycosyltransferase TGF-β1 コア2合成酵素

项目摘要

Core2 glycosyltransferase (C2GnT) is a key enzyme in the synthesis of polylactosamine structure and sialyl Lewis x or a determinant, As these determinants are ligands for adhesion molecules, this enzyme plays an important role in cancer metastasis and inflammation. The aim in this experiment is to examine the effect of various cytokines on the activity of C2GnT in Caco-2 cells.Material and Method :1.Activity of C2GnT in the presence or absence of various cytokines such as TGF-beta1, EGF, basic FGF, HGP2.Time course study of C2GnT activity in the presence of 5ng/ml of TGF-beta1 which gave the highest C2Gn Tactivity3.Expression of C2GnT mRNA under various concentrations of TGF-beta1 (RT-PCR)4. Expression of TGF-beta receptors in Caco-2 cells (immunostaining)5.Effect ofTGF-beta1 on cell growth (MTT)Results :1.Activity of C2GnT increased in proportion to the concentrations of TGF-beta1(1*10 ng/ml).5ng/ml of TGF-beta1 gave the highest C2GnT activity.No increase was observed by the addition of EGF, basic FGF or HGF.2.Increased activity was observed 24 hours after the treatment until 72 hours.3.Increased expression of C2GnT mRNA was observed by the addition of 1, 5, 10 ng/ml of TGF-beta1.The strongest expression was observed at the concentration of 10ng/ ml.4.Both receptors were present on the surface of Caco-2 cells.5.TGF-beta1 suppressed the growth of Caco-2 cells in a dose-dependent manner.Smmary : TGF-beta1 increases the activity of Core2 glycosyltransferase by up-regulating the expression of C2GnT mRNA.Previous reports suggest the relationship between increased C2GnT activity and invasion of colon cancer. We are going to examine whether antisense oligonucleotides against C2GnT suppress the Invasion of colon cancer cells in vitro.

Core2糖基转移酶（C2GnT）是合成聚乳糖胺结构和唾液酸Lewis x或决定簇的关键酶，由于这些决定簇是粘附分子的配体，因此该酶在癌症转移和炎症中发挥重要作用。本实验的目的是检查Caco-2细胞中各种细胞因子对C2GnT活性的影响。材料和方法：1.在存在或不存在各种细胞因子（例如TGF-β1、EGF、碱性FGF、HGP2）的情况下C2GnT的活性。在5ng/ml的TGF-β1存在下C2GnT活性的时程研究，其给出最高的C2Gn Tactivity3.不同浓度TGF-beta1下C2GnT mRNA的表达(RT-PCR)4。 Caco-2细胞中TGF-β受体的表达（免疫染色）5.TGF-β1对细胞生长的影响（MTT）结果：1.C2GnT活性与TGF-β1浓度成比例增加（1*10ng/ml）。5ng/ml TGF-β1给出最高的C2GnT活性。添加EGF、碱性后未观察到增加。 FGF或HGF。2.处理后24小时直至72小时观察到活性增加。3.添加1、5、10ng/ml的TGF-β1观察到C2GnT mRNA表达增加。在10ng/ml浓度时观察到最强表达。4.两种受体均存在于Caco-2细胞表面。5.TGF-β1抑制FGF或HGF的生长。 Caco-2细胞 Smmary：TGF-β1通过上调C2GnT mRNA的表达来增加Core2糖基转移酶的活性。之前的报道表明C2GnT活性增加与结肠癌侵袭之间存在关系。我们将在体外研究针对 C2GnT 的反义寡核苷酸是否抑制结肠癌细胞的侵袭。