Activation of a beta 2-adrenergic receptor/Gs alpha fusion protein elicits a desensitization-resistant cAMP signal capable of inhibiting proliferation of two cancer cell lines.
We showed in a previous study that the expression, in Gs-deficient S49 cyc- cells, of a fusion gene encoding the beta 2-adrenergic Receptor (beta 2AR) and the alpha subunit of the Gs protein (Gs alpha) restored beta 2AR-dependent activation of adenylyl cyclase. We report here the extensive characterization of short- and long-term regulation of the beta 2AR/Gs alpha fusion protein activity and its pharmacological effect after expression in two cancer cell lines. In contrast with native beta 2ARs and Gs, the receptor and the alpha s subunit moieties of the beta 2AR/Gs alpha fusion protein did not undergo functional uncoupling. After a sustained incubation with isoproterenol or forskolin, the accumulation of cAMP could still be observed in S49 beta Gs cells, expressing the fusion gene, which showed, in addition, an up-regulation of their beta 2AR binding sites, while in S49 wt cells, the same treatments completely abolished the rise of cAMP and markedly reduced the number of receptors. cAMP-activation of protein kinase A (PKA) is known to modulate proliferation of most cells. We studied the effect of long term beta 2AR/Gs alpha activation on the growth rate of S49 lymphoma cells and carcinoma carB cells, a highly proliferative cancer cell line expressing oncogenic ras protein. The beta 2AR agonist salmeterol blocked the proliferation of both S49 and carB beta 2Gs cells, while this treatment did not change the growth of wild-type cells. In carB beta 2Gs cells, this effect may be reinforced by a significant basal activity of the fusion protein and by agonist-promoted MAP kinase inhibition. In conclusion, the stimulatory overload provided by the beta 2AR/Gs alpha fusion protein led to the inhibition of cAMP-sensitive cancer cell proliferation in vitro.
Bertin
B
B
Institut Cochin de Génétique Moléculaire, CNRS UPR 415 Paris, France.
Jockers
R
R
Strosberg
A D
AD
Marullo
S
S
D000262:Adenylyl Cyclases / Q000378:metabolism; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000420:Albuterol / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002455:Cell Division; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D015536:Down-Regulation; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D020120:Iodocyanopindolol; D007545:Isoproterenol / Q000494:pharmacology; D051379:Mice; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D000068299:Salmeterol Xinafoate; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured; D018631:ras Proteins / Q000235:genetics
D000318:Adrenergic beta-Agonists; D018343:Receptors, Adrenergic, beta-2; D011993:Recombinant Fusion Proteins; D005576:Colforsin; D000068299:Salmeterol Xinafoate; D020120:Iodocyanopindolol; D010869:Pindolol; D000242:Cyclic AMP; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019205:GTP-Binding Protein alpha Subunits, Gs; D018631:ras Proteins; D000262:Adenylyl Cyclases; D007545:Isoproterenol; D000420:Albuterol