CholecystokininB Receptor from Human Jurkat Lymphoblastic T Cells Is Involved in Activator Protein-1-Responsive Gene Activation

  1. Catherine Oiry1,
  2. Didier Gagne1,
  3. Éric Cottin,
  4. Nicole Bernad1,
  5. Jean-Claude Galleyrand1,
  6. Gilbert Bergé1,
  7. Marie-Francoise Lignon1,
  8. Patrick Eldin2,
  9. Martine Le Cunff2,
  10. Jean Léger2,
  11. Pascal Clerc3,
  12. Daniel Fourmy3 and
  13. Jean Martinez1
  1. 1Laboratoire des Amino Acides, Peptides et Protéines (L.A.P.P.) ESA.CNRS 5075, Universités de Montpellier I et II Faculté de Pharmacie, 34060 Montpellier, France (C.O., D.G., E.C., N.B., J.-C.G., G.B., M.-F.L., J.M.), 2Institut National de la Santé et de la Recherche Médicale U300, Faculté de Pharmacie, 34060 Montpellier, France (P.E., M. le C., J.L.), and3Institut National de la Santé et de la Recherche Médicale U151, Centre Hospitalier Universitaire Rangueil, 31054 Toulouse, France (P.C., D.F.)

    Abstract

    The aim of this study was to analyze the role of cholecystokinin (CCKB) receptor in human lymphoblastic Jurkat T cells. We investigated the trophic effect resulting from activation of such a receptor by using the reporter gene strategy. For this purpose, we transiently transfected Jurkat T cells with the reporter plasmid p[(TRE)3-tk-Luc] and found that CCK-8 was able to dose-dependently induce luciferase expression related to activator protein-1 (AP-1) activation with a maximal response identical to that obtained with compounds known to activate AP-1 complex (quantitatively, the same level of induction was obtained with 1 nm12-O-tetradecanoylphorbol-13-acetate, 100 μm diacylglycerol, or 4 nm epidermal growth factor). The involvement of the CCKB receptor in such a stimulation was demonstrated by the inhibiting effect of the selective CCKB receptor antagonist PD-135,158. This effect was confirmed in COS-7 cells transfected with the cDNA of CCKBreceptor cloned from Jurkat T cells. To better understand the AP-1-dependent luciferase expression in Jurkat T cells, we tested two specific inhibitors of serine/threonine phosphatases-1 and -2A: okadaic acid and calyculin A. These compounds strongly increased the phorbol-12-myristate-13-acetate response, whereas we have not observed a contribution of phosphatase inhibitors on a CCK-8-induced luciferase activity. To confirm that CCKB receptors are involved in AP-1 response, we investigated the CCK-8 effect on interleukin-2 expression, a natural endogenous gene regulated by several factors, including AP-1. In Jurkat T cells activated by phorbol-12-myristate-13-acetate and phytohemagglutinin, CCK-8 induced IL-2 expression. This induction was abolished by PD-135,158. Our results indicate that CCK-8 exerts a trophic effect in Jurkat T cells through stimulation of CCKB receptors by modulation of expression of AP-1-regulated genes.

    Footnotes

    • Send reprint requests to: Jean Martinez, Director, Laboratoire des Amino Acides, Peptides et Protéines (L.A.P.P.), CNRS-ESA 5075, Universités Montpellier I et II, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier Cedex, France.

    • ↵1 M.-F. Lignon and J. Martinez, unpublished observations.

    • ↵2 D. Gagne and J. Martinez, unpublished observations.

    • This work was supported in part by Grant 6846 from Association pour la Recherche contre le Cancer (ARC), Villejuif, France.

    • Abbreviations:
      CCK
      cholecystokinin
      CCK-8
      H-Asp-Tyr(SO3H)-Met-Gly-Trp-Met-Asp-Phe-NH2
      125I-BH-CCK-8
      Bolton-Hunter cholecystokinin(26–33)
      gastrin(5–17)
      H-Leu-(Glu)5-Ala-Tyr-Gly-Trp-Met-Asp-Phe-NH2
      BOC-CCK-4
      N-tert-butoxycarbonyl-Trp-Met-Asp-Phe-NH2
      YM-022
      (R)-1-[2,3-dihydro-1-(2′-methyl-phenacyl)-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl]-3-(3-methylphenyl)urea
      PD-135
      158 [4([2-[[3-(1H-indol-3-yl)-2-methyl-1-oxo-2-[[[1.7.7-trimethyl-bicyclo[2.2.1]hept-2-yl)oxy]carbonyl]amino]propyl]amino]1-phenylethyl]amino-4-oxo-[1S1α.,2β[S*(S*)]4α]]butanoateN-methyl-d-glucamine (bicyclo system 1S-endo)
      L-365
      260, (3R)-(+)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N-(3-methyl-phenyl)urea
      L-364
      718, (3S)-(−)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-1H-indole-2-carboximide
      AP-1
      activator protein-1
      TRE
      12-O-tetradecanoylphorbol-13-acetate-responsive element
      Luc
      luciferase
      PKC
      protein kinase C
      PMA
      phorbol-12-myristate-13-acetate
      PHA
      phytohemagglutinin
      FCS
      fetal calf serum
      PCR
      polymerase chain reaction
      IL-2
      interleukin-2
      HEPES
      4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
      PP-1
      protein phosphatase-1
      PP-2A
      protein phosphatase-2A
      • Received January 27, 1997.
      • Accepted April 28, 1997.
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    1. Molecular Pharmacology August 1, 1997 vol. 52 no. 2 292-299
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