title: Expression of a splice variant of the receptor for GHRH in 3T3 fibroblasts activates cell proliferation responses to GHRH analogs.
abstract: The stimulatory effects of growth hormone-releasing hormone (GHRH) and the antiproliferative action of GHRH antagonists have been demonstrated in various cancers, but the receptors that mediate these responses are not clearly identified. Recently, we reported that human cancer cell lines express splice variants (SVs) of the receptors for GHRH. SV1 exhibits the greatest similarity to the pituitary GHRH receptor and is most likely to be functional. To ascertain whether SV1 mediates mitogenic effects on nonpituitary tissues, we expressed SV1 in 3T3 mouse fibroblasts and studied the properties of the transfected cells. Radioligand binding assays with (125)I-labeled GHRH antagonist JV-1-42 detected high affinity (K(d) = 0.58 +/- 0.17 nM) binding sites for GHRH with a maximal binding capacity (B(max)) of 103 +/- 17.4 fmol/mg of membrane protein in 3T3 cells transfected with pcDNA3-SV1, whereas the control cells transfected with the empty vector did not show any GHRH binding. Cell proliferation studies showed that cells expressing SV1 are much more sensitive to GHRH analogs than the pcDNA3 controls. Thus, the expression of SV1 augments the stimulatory responses to GHRH(1-29)NH(2) or GHRH agonist JI-38 and inhibitory responses to GHRH antagonist JV-1-38 as compared with pcDNA3 controls. The stimulation of SV1-expressing cells by GHRH or JI-38 is followed by an increase in cAMP production, but no GH release occurs. Vasoactive intestinal peptide had no effect, and its antagonist JV-1-53 did not inhibit the proliferation of SV1-expressing cells stimulated by GHRH. Our results suggest that SV1 could mediate responses of nonpituitary cells and various tumors to GHRH and GHRH antagonists. The presence of SV1 in several human cancer cell lines provides a rationale for antitumor therapy based on the blockade of this receptor by specific GHRH antagonists.
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA.
authors.forename: Hippokratis
authors.initials: H
authors.lastname: Kiaris
authors.forename: Andrew V
authors.initials: AV
authors.lastname: Schally
authors.forename: Rebeca
authors.initials: R
authors.lastname: Busto
authors.forename: Gabor
authors.initials: G
authors.lastname: Halmos
authors.forename: Spyros
authors.initials: S
authors.lastname: Artavanis-Tsakonas
authors.forename: Jozsef L
authors.initials: JL
authors.lastname: Varga
mesh_terms: D016475:3T3 Cells; D017398:Alternative Splicing; D000818:Animals; D002455:Cell Division; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000378:metabolism; D013006:Growth Hormone / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D051379:Mice; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D018013:Receptors, Neuropeptide / Q000096:biosynthesis* / Q000235:genetics*; D018017:Receptors, Pituitary Hormone-Regulating Hormone / Q000096:biosynthesis* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology
chemical_list: D008024:Ligands; D010455:Peptides; D018013:Receptors, Neuropeptide; D018017:Receptors, Pituitary Hormone-Regulating Hormone; D014660:Vasoactive Intestinal Peptide; D013006:Growth Hormone; D000242:Cyclic AMP; C048250:somatotropin releasing hormone receptor