Pepsinogen C expression in intestinal IEC-6 cells.
Pepsinogen C (PGC) is the inactive precursor of pepsin C, which is a member of the aspartic proteinase family of proteolytic enzymes, and is found within the vertebrate stomach. The precursor molecule is synthesised within the gastric epithelial cells and secreted into the gastric lumen where it undergoes an autocatalytic activation under acidic conditions to the proteolytic molecule. However, the synthesis of PGC has also been reported in numerous non-gastric tissues including the prostate gland and the Brunner's gland of the small intestine. The physiological significance of PGC in these tissues is not known and studies are limited by the lack of appropriate in vitro cell models. We report here the use of the rat intestinal cell line, IEC-6, as an in vitro model to study the role of pepsinogen C in the intestine. PGC expression was detected in the IEC-6 cells by RT-PCR and immunocytochemistry, using a PGC specific antibody, localised the protein to cytoplasmic secretory granules. Enzymatic assay confirmed the presence of a functional protein exhibiting pepsin activity. Using semi-quantitative RT-PCR we showed that PGC gene expression was up-regulated by the gastro-intestinal hormones gastrin and secretin, and forskolin which stimulates adenylate cyclase activity. This study demonstrates that the IEC-6 cells provide a unique in vitro model for studying pepsinogen C and its potential role(s) in the small intestine.
School of Cell Molecular Biosciences, University of Newcastle upon Tyne, United Kingdom.
Michelle
M
Wells
Brian
B
Brown
Judith
J
Hall
D000818:Animals; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D005755:Gastrins / Q000494:pharmacology; D007421:Intestine, Small / Q000166:cytology / Q000201:enzymology*; D020115:Pepsinogen C / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D012633:Secretin / Q000494:pharmacology
D005755:Gastrins; D012333:RNA, Messenger; D012633:Secretin; D005576:Colforsin; D020115:Pepsinogen C