Regulatory mechanism of pepsinogen gene expression in monolayer cultured guinea pig gastric chief cells

• Published in: Digestive diseases and sciences Vol. 43; no. 12; pp. 2744 - 2749
• Main Authors: OKAYAMA, N, ITOH, M, JOH, T, MIYAMOTO, T, TAKEUCHI, T, MORIYAMA, A, KATO, T
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 01.12.1998; Springer Nature B.V
• Subjects: Animals; Biological and medical sciences; Carbachol - pharmacology; Cells, Cultured; Chief Cells, Gastric - metabolism; Colforsin - pharmacology; Cyclic AMP - pharmacology; Electrophoresis, Capillary; Fundamental and applied biological sciences. Psychology; Gastrointestinal hormones; Gene Expression Regulation - drug effects; Guinea Pigs; Ionomycin - pharmacology; Male; Pepsinogens - genetics; Pepsinogens - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - biosynthesis; Tetradecanoylphorbol Acetate - pharmacology; Up-Regulation; Vertebrates: digestive system; Cell culture; Stomach; Calcium; Digestive system; Rodentia; Cyclic AMP; Experimental study; Gene expression; Pepsinogen; Vertebrata; Regulation(control); Mammalia; Animal; Biological effect; Intracellular; Endocrinology; Hamster
• ISSN: 0163-2116; 1573-2568
• DOI: 10.1023/A:1026623932350

We have investigated the effects of dibutyryl cAMP, forskolin, carbamylcholine chloride (carbachol), ionomycin, and 12-O-tetradecanoylphorbol-13-acetate (TPA) on the expression of guinea pig pepsinogen mRNA in monolayer cultured gastric chief cells. After exposure of the cells to each of these compounds for 4 to 24 hr, and at 48 hr after primary culture, total cellular RNA was isolated using acid guanidium-phenol-chloroform and then was reverse transcribed to cDNA. Obtained cDNA was amplified by polymerase chain reaction (PCR) using primers detecting guinea pig pepsinogen mRNA and human beta-actin mRNA as an internal standard. The PCR products were separated and quantified using capillary electrophoresis. Dibutyryl cAMP and forskolin significantly increased pepsinogen mRNA, but carbachol, ionomycin, and TPA failed to increase that. These findings suggested that pepsinogen gene expression was up-regulated by intracellular cAMP, but not by intracellular calcium or protein kinase C in guinea pig chief cells.