The effect of various in flammatory cytokines in Core2 glycosyltransferase
Grant-in-Aid for Scientific Research (C)
|Research Institution||Second department of Internal medicine, Osaka Medical college|
MAEMURA Kentarou OSAKA MEDICAL COLLEGE,MEDICINE,ASSISTANT, 医学部, 助手 (50281506)
熊野 宏二 大阪医科大学, 医学部, 助手
KUMANO Kouji OSAKA MEDICAL COLLEGE,MEDICINE
|Project Fiscal Year
1997 – 1998
Completed(Fiscal Year 1998)
|Budget Amount *help
¥1,700,000 (Direct Cost : ¥1,700,000)
Fiscal Year 1998 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1997 : ¥900,000 (Direct Cost : ¥900,000)
|Keywords||TGF-beta / Core2 glycosyltransferase / TGF-β / コア2転移酵素 / TGF-β1 / コア2合成酵素|
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, HGP
2.Time course study of C2GnT activity in the presence of 5ng/ml of TGF-beta1 which gave the highest C2Gn Tactivity
3.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)
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.
Research Output (3results)