The CCK-2/gastrin splice variant receptor retaining intron 4 transactivates the COX-2 promoter in vitro.

The CCK-2/gastrin splice variant receptor retaining intron 4 transactivates the COX-2 promoter in vitro. Research Abstract Details 

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The CCK-2/gastrin splice variant receptor retaining intron 4 transactivates the COX-2 promoter in vitro. Abstract Text:

He Huang,Nikolaus Ansorge,Henning Schrader,Matthias Banasch,Hong-Gang Yu,Wolfgang E Schmidt,Michael ,Frank Schmitz,He Huang,Nikolaus Ansorge,Henning Schrader,Matthias Banasch,Hong-Gang Yu,Wolfgang E Schmidt,Michael ,Frank Schmitz,

The expression of the human cholecystokinin-2/gastrin receptor (CCK-2R) has been widely reported in human colorectal cancers. Recently, a splice variant of the CCK-2R retaining intron 4 (CCK-2i4svR) has been cloned from human colorectal cancers and postulated to exhibit constitutive activity. But its role in mediating carcinogenic effects of mature-amidated gastrin in colorectal cancers has not been fully explored. The purpose of the present study was to determine whether the activation of CCK-2i4svR by gastrin transactivates the COX-2 promoter in human colon cancer cells and in COS-7 cells. In this study, Colo320 cells and COS-7 cells were transfected with the human CCK-2R wild type (CCK-2wtR) (COS-7WT, Colo320WT) and the human CCK-2i4svR (COS-7SV, Colo320SV) cDNA. After stimulation with gastrin-17 (G-17), transactivation of the COX-2 promoter was determined by luciferase reporter gene assay. 5'deletions of the COX-2 promoter were transfected into Colo320 cells to narrow down the minimally required regulatory element. Induction of COX-2 expression was further explored at the mRNA level using real time RT-PCR. The effects of CCK-2i4svR activation on phosphorylation of ERK1/2, p38(MAPK) and JNK were examined by using immunoblotting. Prostaglandin E(2) (PGE(2)) secretion was measured by ELISA. Our results showed that gastrin transactivates the COX-2 promoter in both Colo320 cells and COS-7 cells expressing the CCK-2i4svR cDNA. Inhibition of p38(MAPK) pathway using specific inhibitor significantly blocked the gastrin-induced COX-2 transactivation. Gastrin time-dependently increased COX-2 mRNA expression, the peak mRNA levels appeared at 10 h after stimulation. PGE(2) secretion from gastrin-treated cells increased significantly 8 h after stimulation. Treatment with gastrin also stimulated PGE(2) secretion in the Colo320 cells expressing CCK-2i4svR. In conclusion, the CCK-2i4svR mediates transactivation of the COX-2 promoter and MAPK pathway is involved in this process.

The CCK-2/gastrin splice variant receptor retaining intron 4 transactivates the COX-2 promoter in vitro. Publishing Authors By Initials

H Huang,N Ansorge,H Schrader,M Banasch,HG Yu,WE Schmidt,M ,F Schmitz,H Huang,N Ansorge,H Schrader,M Banasch,HG Yu,WE Schmidt,M ,F Schmitz,

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The CCK-2/gastrin splice variant receptor retaining intron 4 transactivates the COX-2 promoter in vitro. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Regulatory peptides

VOLUME: 144

Page Numbers: 34-42

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ISSN: 0167-0115

DAY: 7

MONTH: 06

YEAR: 2007

The CCK-2/gastrin splice variant receptor retaining intron 4 transactivates the COX-2 promoter in vitro. Information

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LANGUAGE: eng

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Country: Netherlands

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