A Functional in Vitro Model to Examine Signaling Mechanisms in Gastrin-Mediated Human Cell Growth

Hong Jin Kim, B. Mark Evers, Nitesh A. Banker, Mark R. Hellmich, Courtney M. Towsend

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Gastrin (G-17) is a trophic hormone with a high affinity for the cholecystokinin-B receptor (CCK-BR); the mechanisms linking receptor binding and activation of downstream events to cell growth are not known, and these studies have been hampered by the lack of a cell model. We have established a pancreatic carcinoid cell line, BON, which produces a number of gut hormones; however, these cells lack native CCK-BR. The purpose of our study was to develop a model cell line containing the CCK-BR and to characterize the cellular mechanisms involved in gastrin regulation of human cell growth. BON cells were transfected with an expression plasmid containing the human CCK-BR, and stable clones were selected using G418. Functional CCK-BR was confirmed by reverse transcriptase-polymerase chain reaction, 125 I-gastrin binding, and mobilization of intracellular calcium ([Ca 2+ ] i ) in response to G-17. Stable transfectants were treated with G-17 (±) the CCK-BR antagonist, L365,260 (L-60); growth was assessed using a Coulter counter. G-17 stimulated the growth of the stable clones, whereas the selective CCK-BR antagonist, L-60, abolished this G-17-mediated trophic effect. We have shown that G-17, acting through the CCK-BR, mobilizes [Ca 2+ ] i as a second messenger and stimulates cell growth. Our unique BON cell line, stably transfected with the human CCK-BR, provides a novel paradigm to further delineate signaling mechanisms in gastrin regulation of human cell growth.

Original languageEnglish (US)
Pages (from-to)69-77
Number of pages9
JournalJournal of Gastrointestinal Surgery
Volume1
Issue number1
DOIs
StatePublished - Jan 1 1997

ASJC Scopus subject areas

  • Surgery
  • Gastroenterology

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