The effect of various in flammatory cytokines in Core2 glycosyltransferase

Core2糖基转移酶（Core2 glycosyltransferase, C2GnT）是合成聚乳糖胺结构和唾液酰基Lewis x或一种决定因子的关键酶，这些决定因子是粘附分子的配体，在肿瘤转移和炎症反应中起重要作用。本实验旨在探讨不同细胞因子对Caco-2细胞C2GnT活性的影响。材料与方法：C2GnT在各种细胞因子如tgf - β 1、EGF、碱性FGF、HGP2存在或不存在时的活性。5ng/ml tgf - β存在时C2GnT活性的时间过程研究，其中5ng/ml的tgf - β具有最高的C2GnT活性3。不同浓度tgf - β 1 （RT-PCR）下C2GnT mRNA的表达tgf - β受体在Caco-2细胞中的表达（免疫染色）tgf - β 1对细胞生长的影响（MTT）C2GnT活性随tgf - β 1浓度（1*10 ng/ml）成比例增加。5ng/ml的tgf - β 1具有最高的C2GnT活性。添加EGF、碱性FGF或HGF.2均未见升高。治疗后24小时至72小时观察到活性增加。添加1、5、10 ng/ml tgf - β 1后，C2GnT mRNA表达增加。在浓度为10ng/ ml时表达最强。这两种受体均存在于Caco-2细胞表面。tgf - β 1以剂量依赖性方式抑制Caco-2细胞的生长。摘要：tgf - β 1通过上调C2GnT mRNA的表达来增加Core2糖基转移酶的活性。先前的报告表明，C2GnT活性增加与结肠癌侵袭之间存在关系。我们将在体外研究抗C2GnT的反义寡核苷酸是否抑制结肠癌细胞的侵袭。

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.