Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1997 Jun 1;99(11):2762–2771. doi: 10.1172/JCI119466

EGF receptor activation stimulates endogenous gastrin gene expression in canine G cells and human gastric cell cultures.

M G Ford 1, J D Valle 1, C J Soroka 1, J L Merchant 1
PMCID: PMC508123  PMID: 9169507

Abstract

Gastrin release from the antral gastrin-expressing cell (G cell) is regulated by bombesin and luminal factors. Yet, these same extracellular regulators do not stimulate expression of the gene. Since the gastric mucosa expresses large quantities of EGF receptor ligands such as TGFalpha, we examined whether EGF receptor ligands stimulate gastrin gene expression in gastrin-expressing cell cultures. EGF receptor activation of primary cultures stimulated gastrin gene expression about twofold; whereas bombesin treatment of antral G cell cultures stimulated gastrin release but not gene expression. EGF and TGFalpha were weak stimulants of gastrin release. EGF receptor activation of AGS human gastric adenocarcinoma cell line stimulated gastrin gene expression nearly fourfold; and gastrin reporter constructs transfected into AGS cells were stimulated more than fourfold by EGF. EGF induction was conferred by the previously defined GC-rich gastrin EGF response element (gERE) element located at -68 to -53 bp upstream from the cap site since a mutation of the gERE element abolished both basal and EGF induction. Moreover, EGF treatment of AGS cells stimulated binding of the transcription factor Sp1 to this element. Collectively, these results demonstrate that gastrin gene expression and gastrin release are regulated by different signaling pathways: gene expression by EGF receptor activation and gastrin secretion by neuropeptides and luminal factors.

Full Text

The Full Text of this article is available as a PDF (924.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bandyopadhyay G. K., Imagawa W., Wallace D. R., Nandi S. Proliferative effects of insulin and epidermal growth factor on mouse mammary epithelial cells in primary culture. Enhancement by hydroxyeicosatetraenoic acids and synergism with prostaglandin E2. J Biol Chem. 1988 Jun 5;263(16):7567–7573. [PubMed] [Google Scholar]
  2. Barranco S. C., Townsend C. M., Jr, Casartelli C., Macik B. G., Burger N. L., Boerwinkle W. R., Gourley W. K. Establishment and characterization of an in vitro model system for human adenocarcinoma of the stomach. Cancer Res. 1983 Apr;43(4):1703–1709. [PubMed] [Google Scholar]
  3. Beauchamp R. D., Barnard J. A., McCutchen C. M., Cherner J. A., Coffey R. J., Jr Localization of transforming growth factor alpha and its receptor in gastric mucosal cells. Implications for a regulatory role in acid secretion and mucosal renewal. J Clin Invest. 1989 Sep;84(3):1017–1023. doi: 10.1172/JCI114223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brand S. J., Stone D. Reciprocal regulation of antral gastrin and somatostatin gene expression by omeprazole-induced achlorhydria. J Clin Invest. 1988 Sep;82(3):1059–1066. doi: 10.1172/JCI113662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bundgaard J. R., Hansen T. O., Friis-Hansen L., Rourke I. J., van Solinge W. W., Nielsen F. C., Rehfeld J. F. A distal Sp1-element is necessary for maximal activity of the human gastrin gene promoter. FEBS Lett. 1995 Aug 7;369(2-3):225–228. doi: 10.1016/0014-5793(95)00754-w. [DOI] [PubMed] [Google Scholar]
  6. Campos R. V., Buchan A. M., Meloche R. M., Pederson R. A., Kwok Y. N., Coy D. H. Gastrin secretion from human antral G cells in culture. Gastroenterology. 1990 Jul;99(1):36–44. doi: 10.1016/0016-5085(90)91226-v. [DOI] [PubMed] [Google Scholar]
  7. DelValle J., Yamada T. Amino acids and amines stimulate gastrin release from canine antral G-cells via different pathways. J Clin Invest. 1990 Jan;85(1):139–143. doi: 10.1172/JCI114404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dempsey P. J., Goldenring J. R., Soroka C. J., Modlin I. M., McClure R. W., Lind C. D., Ahlquist D. A., Pittelkow M. R., Lee D. C., Sandgren E. P. Possible role of transforming growth factor alpha in the pathogenesis of Ménétrier's disease: supportive evidence form humans and transgenic mice. Gastroenterology. 1992 Dec;103(6):1950–1963. doi: 10.1016/0016-5085(92)91455-d. [DOI] [PubMed] [Google Scholar]
  9. Deng W. P., Nickoloff J. A. Site-directed mutagenesis of virtually any plasmid by eliminating a unique site. Anal Biochem. 1992 Jan;200(1):81–88. doi: 10.1016/0003-2697(92)90280-k. [DOI] [PubMed] [Google Scholar]
  10. Gantz I., Takeuchi T., Yamada T. Cloning of canine gastrin cDNA's encoding variant amino acid sequences. Digestion. 1990;46 (Suppl 2):99–104. doi: 10.1159/000200372. [DOI] [PubMed] [Google Scholar]
  11. Giraud A. S., Soll A. H., Cuttitta F., Walsh J. H. Bombesin stimulation of gastrin release from canine gastrin cells in primary culture. Am J Physiol. 1987 Mar;252(3 Pt 1):G413–G420. doi: 10.1152/ajpgi.1987.252.3.G413. [DOI] [PubMed] [Google Scholar]
  12. Godley J. M., Brand S. J. Regulation of the gastrin promoter by epidermal growth factor and neuropeptides. Proc Natl Acad Sci U S A. 1989 May;86(9):3036–3040. doi: 10.1073/pnas.86.9.3036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goodlad R. A., Ghatei M. A., Domin J., Bloom S. R., Gregory H., Wright N. A. Plasma enteroglucagon, peptide YY and gastrin in rats deprived of luminal nutrition, and after urogastrone-EGF administration. A proliferative role for PYY in the intestinal epithelium? Experientia. 1989 Feb 15;45(2):168–169. doi: 10.1007/BF01954862. [DOI] [PubMed] [Google Scholar]
  14. Heitz P. U., Kasper M., van Noorden S., Polak J. M., Gregory H., Pearse A. G. Immunohistochemical localisation of urogastrone to human duodenal and submandibular glands. Gut. 1978 May;19(5):408–413. doi: 10.1136/gut.19.5.408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Howell G. M., Ziober B. L., Humphrey L. E., Willson J. K., Sun L., Lynch M., Brattain M. G. Regulation of autocrine gastrin expression by the TGF alpha autocrine loop. J Cell Physiol. 1995 Feb;162(2):256–265. doi: 10.1002/jcp.1041620211. [DOI] [PubMed] [Google Scholar]
  16. Ikeda H., Mitsuhashi T., Kubota K., Kuzuya N., Uchimura H. Epidermal growth factor stimulates growth hormone secretion from superfused rat adenohypophyseal fragments. Endocrinology. 1984 Aug;115(2):556–558. doi: 10.1210/endo-115-2-556. [DOI] [PubMed] [Google Scholar]
  17. Johnson L. K., Baxter J. D., Vlodavsky I., Gospodarowicz D. Epidermal growth factor and expression of specific genes: effects on cultured rat pituitary cells are dissociable from the mitogenic response. Proc Natl Acad Sci U S A. 1980 Jan;77(1):394–398. doi: 10.1073/pnas.77.1.394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaise M., Muraoka A., Yamada J., Yamada T. Epidermal growth factor induces H+,K+-ATPase alpha-subunit gene expression through an element homologous to the 3' half-site of the c-fos serum response element. J Biol Chem. 1995 Aug 4;270(31):18637–18642. doi: 10.1074/jbc.270.31.18637. [DOI] [PubMed] [Google Scholar]
  19. Koh T. J., Wang T. C. Molecular cloning and sequencing of the murine gastrin gene. Biochem Biophys Res Commun. 1995 Nov 2;216(1):34–41. doi: 10.1006/bbrc.1995.2588. [DOI] [PubMed] [Google Scholar]
  20. Logsdon C. D., Moessner J., Williams J. A., Goldfine I. D. Glucocorticoids increase amylase mRNA levels, secretory organelles, and secretion in pancreatic acinar AR42J cells. J Cell Biol. 1985 Apr;100(4):1200–1208. doi: 10.1083/jcb.100.4.1200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Logsdon C. D. Stimulation of pancreatic acinar cell growth by CCK, epidermal growth factor, and insulin in vitro. Am J Physiol. 1986 Oct;251(4 Pt 1):G487–G494. doi: 10.1152/ajpgi.1986.251.4.G487. [DOI] [PubMed] [Google Scholar]
  22. Marshak D. W., Aldrich L. B., Del Valle J., Yamada T. Localization of immunoreactive cholecystokinin precursor to amacrine cells and bipolar cells of the macaque monkey retina. J Neurosci. 1990 Sep;10(9):3045–3055. doi: 10.1523/JNEUROSCI.10-09-03045.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Massagué J. Epidermal growth factor-like transforming growth factor. II. Interaction with epidermal growth factor receptors in human placenta membranes and A431 cells. J Biol Chem. 1983 Nov 25;258(22):13614–13620. [PubMed] [Google Scholar]
  24. Merchant J. L., Demediuk B., Brand S. J. A GC-rich element confers epidermal growth factor responsiveness to transcription from the gastrin promoter. Mol Cell Biol. 1991 May;11(5):2686–2696. doi: 10.1128/mcb.11.5.2686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Merchant J. L., Iyer G. R., Taylor B. R., Kitchen J. R., Mortensen E. R., Wang Z., Flintoft R. J., Michel J. B., Bassel-Duby R. ZBP-89, a Krüppel-like zinc finger protein, inhibits epidermal growth factor induction of the gastrin promoter. Mol Cell Biol. 1996 Dec;16(12):6644–6653. doi: 10.1128/mcb.16.12.6644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Merchant J. L., Shiotani A., Mortensen E. R., Shumaker D. K., Abraczinskas D. R. Epidermal growth factor stimulation of the human gastrin promoter requires Sp1. J Biol Chem. 1995 Mar 17;270(11):6314–6319. doi: 10.1074/jbc.270.11.6314. [DOI] [PubMed] [Google Scholar]
  27. Murayama Y., Miyagawa J., Higashiyama S., Kondo S., Yabu M., Isozaki K., Kayanoki Y., Kanayama S., Shinomura Y., Taniguchi N. Localization of heparin-binding epidermal growth factor-like growth factor in human gastric mucosa. Gastroenterology. 1995 Oct;109(4):1051–1059. doi: 10.1016/0016-5085(95)90562-6. [DOI] [PubMed] [Google Scholar]
  28. Polk D. H., Ervin M. G., Padbury J. F., Lam R. W., Reviczky A. L., Fisher D. A. Epidermal growth factor acts as a corticotropin-releasing factor in chronically catheterized fetal lambs. J Clin Invest. 1987 Mar;79(3):984–988. doi: 10.1172/JCI112910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Przylipiak A., Kiesel L., Rabe T., Helm K., Przylipiak M., Runnebaum B. Epidermal growth factor stimulates luteinizing hormone and arachidonic acid release in rat pituitary cells. Mol Cell Endocrinol. 1988 May;57(1-2):157–162. doi: 10.1016/0303-7207(88)90045-7. [DOI] [PubMed] [Google Scholar]
  30. Schonbrunn A., Krasnoff M., Westendorf J. M., Tashjian A. H., Jr Epidermal growth factor and thyrotropin-releasing hormone act similarly on a clonal pituitary cell strain. Modulation of hormone production and inhbition of cell proliferation. J Cell Biol. 1980 Jun;85(3):786–797. doi: 10.1083/jcb.85.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schreiber E., Matthias P., Müller M. M., Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. doi: 10.1093/nar/17.15.6419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Searcy R. M., Malagelada J. R. Ménétrièr's disease and idiopathic hypertrophic gastropathy. Ann Intern Med. 1984 Apr;100(4):565–570. doi: 10.7326/0003-4819-100-4-565. [DOI] [PubMed] [Google Scholar]
  33. Shing Y., Christofori G., Hanahan D., Ono Y., Sasada R., Igarashi K., Folkman J. Betacellulin: a mitogen from pancreatic beta cell tumors. Science. 1993 Mar 12;259(5101):1604–1607. doi: 10.1126/science.8456283. [DOI] [PubMed] [Google Scholar]
  34. Shiotani A., Merchant J. L. cAMP regulates gastrin gene expression. Am J Physiol. 1995 Sep;269(3 Pt 1):G458–G464. doi: 10.1152/ajpgi.1995.269.3.G458. [DOI] [PubMed] [Google Scholar]
  35. Singh J. Effects of amino acids, glucagon, insulin and acetylcholine on cyclic nucleotide metabolism and amylase secretion in isolated mouse pancreatic fragments. Biochem Pharmacol. 1983 Jul 1;32(13):2017–2023. doi: 10.1016/0006-2952(83)90421-5. [DOI] [PubMed] [Google Scholar]
  36. Singh J. Mechanism of action of insulin on acetylcholine-evoked amylase secretion in the mouse pancreas. J Physiol. 1985 Jan;358:469–482. doi: 10.1113/jphysiol.1985.sp015562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sugano K., Park J., Soll A. H., Yamada T. Stimulation of gastrin release by bombesin and canine gastrin-releasing peptides. Studies with isolated canine G cells in primary culture. J Clin Invest. 1987 Mar;79(3):935–942. doi: 10.1172/JCI112904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Söling H. D., Unger K. O. The role of insulin in the regulation of -amylase synthesis in the rat pancreas. Eur J Clin Invest. 1972 Jun;2(4):199–212. doi: 10.1111/j.1365-2362.1972.tb00645.x. [DOI] [PubMed] [Google Scholar]
  39. Tahara E., Ito H., Nakagami K., Shimamoto F., Yamamoto M., Sumii K. Scirrhous argyrophil cell carcinoma of the stomach with multiple production of polypeptide hormones, amine, CEA, lysozyme, and HCG. Cancer. 1982 May 1;49(9):1904–1915. doi: 10.1002/1097-0142(19820501)49:9<1904::aid-cncr2820490925>3.0.co;2-i. [DOI] [PubMed] [Google Scholar]
  40. Vinter-Jensen L., Juhl C. O., Rehfeld J. F., Poulsen S. S., Dajani E. Z., Nexø E. Chronic systemic treatment with epidermal growth factor induces hypergastrinaemia in Goettingen minipigs. Scand J Gastroenterol. 1995 May;30(5):422–427. doi: 10.3109/00365529509093301. [DOI] [PubMed] [Google Scholar]
  41. Wang L., Lucey M. R., Fras A. M., Wilson E. J., Del Valle J. Epidermal growth factor and transforming growth factor-alpha directly inhibit parietal cell function through a similar mechanism. J Pharmacol Exp Ther. 1993 Apr;265(1):308–313. [PubMed] [Google Scholar]
  42. Watson S. A., Durrant L. G., Wencyk P. M., Watson A. L., Morris D. L. Intracellular gastrin in human gastrointestinal tumor cells. J Natl Cancer Inst. 1991 Jun 19;83(12):866–871. doi: 10.1093/jnci/83.12.866. [DOI] [PubMed] [Google Scholar]
  43. Wright N. A., Pike C., Elia G. Induction of a novel epidermal growth factor-secreting cell lineage by mucosal ulceration in human gastrointestinal stem cells. Nature. 1990 Jan 4;343(6253):82–85. doi: 10.1038/343082a0. [DOI] [PubMed] [Google Scholar]
  44. Wu S. V., Giraud A., Mogard M., Sumii K., Walsh J. H. Effects of inhibition of gastric secretion on antral gastrin and somatostatin gene expression in rats. Am J Physiol. 1990 May;258(5 Pt 1):G788–G793. doi: 10.1152/ajpgi.1990.258.5.G788. [DOI] [PubMed] [Google Scholar]
  45. Yajima Y., Saito T. The effects of epidermal growth factor on cell proliferation and prolactin production by GH3 rat pituitary cells. J Cell Physiol. 1984 Aug;120(2):249–256. doi: 10.1002/jcp.1041200220. [DOI] [PubMed] [Google Scholar]
  46. Yoshida K., Kyo E., Tsujino T., Sano T., Niimoto M., Tahara E. Expression of epidermal growth factor, transforming growth factor-alpha and their receptor genes in human gastric carcinomas; implication for autocrine growth. Jpn J Cancer Res. 1990 Jan;81(1):43–51. doi: 10.1111/j.1349-7006.1990.tb02505.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Zimmerman R. P., Gates T. S., Boehmer C. G., Mantyh P. W. Epidermal growth factor receptors in the canine antrum. Peptides. 1988 Nov-Dec;9(6):1411–1414. doi: 10.1016/0196-9781(88)90211-2. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES