High and low affinity receptors mediate growth effects of gastrin and gastrin-Gly on DLD-1 human colonic carcinoma cells

Shawn Ahmed, Barna Budai, Krisztina Herédi-Szabó, Judit Farkas, Géza Tóth, Richard F. Murphy, Sándor Lovas

Research output: Contribution to journalArticle

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Abstract

Gastrin (G17) and N-carboxymethylgastrin (G17-Gly) have been shown to stimulate the growth of colon cancer cells both in vivo and in vitro. The identity of the receptor mediating these effects is controversial. A recent study demonstrated the presence of a low affinity binding site for G17 and G17-Gly on the DLD-1 human colon cancer cell line. The goal of the current study was to further investigate the role of this receptor in mediating the growth-promoting effects of gastrin peptides. Binding of [Leu15]G17 and [Leu15]G17-Gly to DLD-1 cell membranes in competition with [ 3H]G17-Gly was examined. Binding of [3H]cholecystokinin-8 (CCK8) to DLD-1 cell membranes was also assessed. Whole cell binding experiments were carried out using [125I-Tyr12,Leu 15]G17-Gly. In addition, the ability of [Leu15]G17 and [Leu15]G17-Gly to stimulate cell growth, as determined by cell counting, was tested. [Leu15]G17 and [Leu15]G17-Gly competed with [3H]G17-Gly at both a high and a low affinity site on DLD-1 membranes. The IC50 values for [Leu15]G17 were 6.0×10-8 M and 6.9×10-6 M while those for [Leu15]G17-Gly were 3.2×10-9 M and 4.9×10 -6 M. [3H]CCK8 did not bind to either site. [Leu 15]G17-Gly also competed with [125I-Tyr 12,Leu15]G17-Gly at both a high and a low affinity site on DLD-1 cells with similar affinities as observed with membranes. [Leu 15]G17 and [Leu15]G17-Gly significantly stimulated the growth of DLD-1 cells in a dose-dependent and biphasic manner. The binding profiles of the peptides tested suggest that these sites are different from previously identified wild-type and mutant CCK1 or CCK2 receptors.

Original languageEnglish
Pages (from-to)199-203
Number of pages5
JournalFEBS Letters
Volume556
Issue number1-3
DOIs
StatePublished - Jan 2 2004

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Gastrins
Cells
Cell membranes
Carcinoma
Growth
Cholecystokinin B Receptor
Membranes
Peptides
Cell growth
Colonic Neoplasms
Binding Sites
Cell Membrane
Inhibitory Concentration 50
Experiments
Cell Line
cholecystokinin 8

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

High and low affinity receptors mediate growth effects of gastrin and gastrin-Gly on DLD-1 human colonic carcinoma cells. / Ahmed, Shawn; Budai, Barna; Herédi-Szabó, Krisztina; Farkas, Judit; Tóth, Géza; Murphy, Richard F.; Lovas, Sándor.

In: FEBS Letters, Vol. 556, No. 1-3, 02.01.2004, p. 199-203.

Research output: Contribution to journalArticle

Ahmed, Shawn ; Budai, Barna ; Herédi-Szabó, Krisztina ; Farkas, Judit ; Tóth, Géza ; Murphy, Richard F. ; Lovas, Sándor. / High and low affinity receptors mediate growth effects of gastrin and gastrin-Gly on DLD-1 human colonic carcinoma cells. In: FEBS Letters. 2004 ; Vol. 556, No. 1-3. pp. 199-203.
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AU - Budai, Barna

AU - Herédi-Szabó, Krisztina

AU - Farkas, Judit

AU - Tóth, Géza

AU - Murphy, Richard F.

AU - Lovas, Sándor

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