A3 adenosine receptor activation in melanoma cells: association between receptor fate and tumor growth inhibition.
Activation of the Gi protein-coupled A3 adenosine receptor (A3AR) has been implicated in the inhibition of melanoma cell growth by deregulating protein kinase A and key components of the Wnt signaling pathway. Receptor activation results in internalization/recycling events that play an important role in turning on/off receptor-mediated signal transduction pathways. Thus, we hereby examined the association between receptor fate, receptor functionality, and tumor growth inhibition upon activation with the agonist 1-deoxy-1-[6-[[(3-iodophenyl)-methyl]amino]-9H-purine-9-yl]-N-methyl-beta-D-ribofuranuronamide (IB-MECA). Results showed that melanoma cells highly expressed A3AR on the cell surface, which was rapidly internalized to the cytosol and "sorted" to the endosomes for recycling and to the lysosomes for degradation. Receptor distribution in the lysosomes was consistent with the down-regulation of receptor protein expression and was followed by mRNA and protein resynthesis. At each stage, receptor functionality was evidenced by the modulation in cAMP level and the downstream effectors protein kinase A, glycogen synthase kinase-3beta, c-Myc, and cyclin D1. The A3AR antagonist MRS 1523 counteracted the internalization process as well as the modulation in the expression of the signaling proteins, demonstrating that the responses are A3AR-mediated. Supporting this notion are the in vivo studies showing tumor growth inhibition upon IB-MECA treatment and reverse of this response when IB-MECA was given in combination with MRS 1523. In addition, in melanoma tumor lesions derived from IB-MECA-treated mice, the expression level A3AR and the downstream key signaling proteins were modulated in the same pattern as was seen in vitro. Altogether, our observations tie the fate of A3AR to modulation of downstream molecular mechanisms leading to tumor growth inhibition both in vitro and in vivo.
Madi
Lea
L
Can-Fite BioPharma Ltd., Kiryat-Matalon, Petach-Tikva 49170, Israel.
Bar-Yehuda
Sara
S
Barer
Faina
F
Ardon
Eti
E
Ochaion
Avivit
A
Fishman
Pnina
P
D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002455:Cell Division; D045744:Cell Line, Tumor; D019938:Cyclin D1 / Q000096:biosynthesis; D015536:Down-Regulation; D008545:Melanoma / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism*; D009374:Neoplasms, Experimental / Q000188:drug therapy; D021381:Protein Transport; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis; D043684:Receptor, Adenosine A3 / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction
D009363:Neoplasm Proteins; D016271:Proto-Oncogene Proteins c-myc; D043684:Receptor, Adenosine A3; D019938:Cyclin D1; C084956:N(6)-(3-iodobenzyl)-5'-N-methylcarboxamidoadenosine; D000241:Adenosine