The effect of various in flammatory cytokines in Core2 glycosyltransferase

Core2糖基转移酶(C2GnT)是合成聚乳糖胺结构和唾液酸化Lewis x或决定簇的关键酶，由于这些决定簇是黏附分子的配体，该酶在肿瘤转移和炎症中起着重要作用。本实验旨在探讨不同细胞因子对Caco-2细胞C2GnT活性的影响。材料和方法：1.在有或无多种细胞因子的作用下，C2GnT的活性。C2GnT活性的时程研究3.不同浓度的转化生长因子-β1对C2GnT mRNA的表达(RT-PCR)4.转化生长因子-β1在Caco-2细胞中的表达(免疫组织化学染色)5.转化生长因子-β1对细胞生长的影响(四甲基偶氮唑盐比色法)结果：1.C2GnT活性与转化生长因子-β1浓度(1*10 ng/ml)成正比，5 ng/ml的转化生长因子-β1对C2GnT活性影响不明显加入1、5、10 ng/ml的转化生长因子-β1后，C2GnT基因表达增强，以10 ng/ml时表达最强。4.两种受体均存在于Caco-2细胞表面。5.转化生长因子-β1呈剂量依赖性抑制Caco-2细胞的生长。我们将检测针对C2GnT的反义寡核苷酸是否在体外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.