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Papers In Press, published online ahead of print June 20, 2002
Gastrointestinal Research, VA Medical Center, San Francisco, CA 94121
Corresponding Author: youngk{at}itsa.ucsf.edu
MUC2 is a secretory mucin normally expressed by goblet cells of the intestinal epithelium. It is over-expressed in mucinous type colorectal cancers but down-regulated in colorectal adenocarcinoma. Phorbol 12-myristate 13-acetate (PMA) treatment of colon cancer cell lines increases MUC2 expression, so we have undertaken a detailed analysis of the effects of PMA on the promoter activity of the 5'-flanking region of the MUC2 gene, using stably and transiently transfected promoter reporter vectors. Protein kinase C (PKC) inhibitors (bisindolylmaleimide, calphostin C) and inhibitors of mitogen-activated protein/extracellular signal regulated kinase kinase (MEK) (PD 98059 and U0126) suppressed up-regulation of MUC2. Src tyrosine kinase inhibitor PP2 , a protein kinase A inhibitor (KT5720), and a p38 inhibitor (SB 203580) did not affect transcription. Western blotting and reverse transcription and polymerase chain reaction (RT-PCR) analysis confirmed these results. In addition, co-transfections with mutants of Ras, Raf and MEK showed that the induction of MUC2 promoter activity by PMA required these three signaling proteins. Our results demonstrate that PMA activates PKC, stimulating MAP kinase through a Ras- and Raf-dependent mechanism. An important role for nuclear factor kB (NF-kB), was also demonstrated using the inhibitor caffeic acid phenethyl ester (CAPE) and electrophoretic mobility shift assays (EMSA). Such identification of pathways involved in MUC2 up-regulation by PMA in the HM3 colon cancer cell line may serve as a model for the effects of cytokines and growth factors which regulate MUC2 expression during the progression of colorectal cancer.
J. Biol. Chem, 10.1074/jbc.M200353200
Submitted on January 11, 2002
Revised on June 20, 2002
Accepted on June 20, 2002
Phorbol 12-myristate 13-acetate up-regulates the transcription of MUC2 intestinal mucin via Ras, ERK and NF-
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