{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T13:59:41Z","timestamp":1766066381727},"reference-count":63,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2014,7,19]],"date-time":"2014-07-19T00:00:00Z","timestamp":1405728000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"FEDER through the Program Operational Competitiveness Factors COMPETE and National Funds through FCT Foundation for Science and Technology","award":["SFRH\/BD\/64911\/2009"],"award-info":[{"award-number":["SFRH\/BD\/64911\/2009"]}]},{"name":"FEDER through the Program Operational Competitiveness Factors COMPETE and National Funds through FCT Foundation for Science and Technology","award":["SFRH\/BPD\/63746\/2009"],"award-info":[{"award-number":["SFRH\/BPD\/63746\/2009"]}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Basic Clin Pharma Tox"],"published-print":{"date-parts":[[2015,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Malignant melanoma is the most deadly type of skin cancer. The lack of effective pharmacological approaches for this tumour can be related to the incomplete understanding of the pathophysiological mechanisms involved in melanoma cell proliferation. Adenosine has growth\u2010promoting and growth inhibitory effects on tumour cells. We aimed to investigate effects of adenosine and its metabolic product, inosine, on human C32 melanoma cells and the signalling pathways involved. The 3\u2010(4,5\u2010dimethylthiazol\u20102\u2010yl)\u20102,5\u2010diphenyl tetrazolium bromide (<jats:styled-content style=\"fixed-case\">MTT<\/jats:styled-content>) reduction and bromodeoxyuridine (BrdU) proliferation assays were used to evaluate adenosine, adenosine deaminase and inosine effects, in the absence or presence of adenosine receptor (<jats:styled-content style=\"fixed-case\">AR<\/jats:styled-content>), A<jats:sub>3<\/jats:sub><jats:styled-content style=\"fixed-case\">AR<\/jats:styled-content> and P2Y<jats:sub>1<\/jats:sub>R antagonists and <jats:styled-content style=\"fixed-case\">PLC<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> PKC<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> MEK<\/jats:styled-content>1\/2 and <jats:styled-content style=\"fixed-case\">PI<\/jats:styled-content>3K inhibitors. <jats:styled-content style=\"fixed-case\">ERK<\/jats:styled-content>1\/2 levels were determined using an <jats:styled-content style=\"fixed-case\">ELISA<\/jats:styled-content> kit. Adenosine and inosine levels were quantified using an enzyme\u2010coupled assay. Adenosine caused cell proliferation through <jats:styled-content style=\"fixed-case\">AR<\/jats:styled-content> activation. Adenosine deaminase increased inosine levels (nanomolar concentrations) on the extracellular space, in a time\u2010dependent manner, inducing proliferation through A<jats:sub>3<\/jats:sub><jats:styled-content style=\"fixed-case\">AR<\/jats:styled-content> activation. Micromolar concentrations of inosine enhanced proliferation through A<jats:sub>3<\/jats:sub><jats:styled-content style=\"fixed-case\">AR<\/jats:styled-content> activation, causing an increase in <jats:styled-content style=\"fixed-case\">ERK<\/jats:styled-content>1\/2 levels, and P2Y<jats:sub>1<\/jats:sub>R activation via <jats:styled-content style=\"fixed-case\">ENT<\/jats:styled-content>\u2010dependent mechanisms. We propose the simultaneous activation of <jats:styled-content style=\"fixed-case\">PLC<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">PKC<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">MEK<\/jats:styled-content>1\/2\u2010<jats:styled-content style=\"fixed-case\">ERK<\/jats:styled-content>1\/2 and <jats:styled-content style=\"fixed-case\">PI<\/jats:styled-content>3K pathways as the main mechanism responsible for the proliferative effect elicited by inosine and its significant role in melanoma cancer progression.<\/jats:p>","DOI":"10.1111\/bcpt.12280","type":"journal-article","created":{"date-parts":[[2014,6,6]],"date-time":"2014-06-06T15:59:09Z","timestamp":1402070349000},"page":"25-36","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Inosine Strongly Enhances Proliferation of Human C32 Melanoma Cells through <scp>PLC<\/scp>\u2010<scp>PKC<\/scp>\u2010<scp>MEK<\/scp>1\/2\u2010<scp>ERK<\/scp>1\/2 and PI3K Pathways"],"prefix":"10.1111","volume":"116","author":[{"given":"Ana Sofia","family":"Soares","sequence":"first","affiliation":[{"name":"REQUIMTE Laborat\u00f3rio de Farmacologia Departamento de Ci\u00eancias do Medicamento Faculdade de Farm\u00e1cia Universidade do Porto Porto Portugal"},{"name":"MedInUP \u2010 Centro de Investiga\u00e7\u00e3o Farmacol\u00f3gica e Inova\u00e7\u00e3o Medicamentosa Universidade do Porto Porto Portugal"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vera Marisa","family":"Costa","sequence":"additional","affiliation":[{"name":"REQUIMTE\/Laborat\u00f3rio de Toxicologia Departamento de Ci\u00eancias Biol\u00f3gicas Faculdade de Farm\u00e1cia Universidade do Porto Porto Portugal"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carmen","family":"Diniz","sequence":"additional","affiliation":[{"name":"REQUIMTE Laborat\u00f3rio de Farmacologia Departamento de Ci\u00eancias do Medicamento Faculdade de Farm\u00e1cia Universidade do Porto Porto Portugal"},{"name":"MedInUP \u2010 Centro de Investiga\u00e7\u00e3o Farmacol\u00f3gica e Inova\u00e7\u00e3o Medicamentosa Universidade do Porto Porto Portugal"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paula","family":"Fresco","sequence":"additional","affiliation":[{"name":"REQUIMTE Laborat\u00f3rio de Farmacologia Departamento de Ci\u00eancias do Medicamento Faculdade de Farm\u00e1cia Universidade do Porto Porto Portugal"},{"name":"MedInUP \u2010 Centro de Investiga\u00e7\u00e3o Farmacol\u00f3gica e Inova\u00e7\u00e3o Medicamentosa Universidade do Porto Porto Portugal"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2014,7,19]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/annonc\/mdp252"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(13)60802-8"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1586\/era.10.170"},{"key":"e_1_2_7_5_1","first-page":"488","article-title":"Treatment of metastatic melanoma: an overview","volume":"23","author":"Bhatia S","year":"2009","journal-title":"Oncology (Williston Park)"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2013.02.056"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.2741\/1755"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1155\/2012\/959848"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11302-013-9372-5"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1155\/2012\/485156"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-94-007-4719-7_2"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10555-007-9044-4"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.5772\/38534"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2003.11.003"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2004.01.006"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2010.292"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.6061\/clinics\/2012(12)15"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.clinbiochem.2005.05.015"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11010-012-1513-6"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2012.08.001"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2013.08.003"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.00708.2012"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bjp.0704349"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1080\/15257770.2011.597372"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.2174\/157016109789043892"},{"key":"e_1_2_7_26_1","doi-asserted-by":"crossref","first-page":"1512","DOI":"10.1182\/blood.V96.4.1512","article-title":"Antileukemic efficacy of 2\u2010deoxycoformycin in monocytic leukemia cells","volume":"96","author":"Niitsu N","year":"2000","journal-title":"Blood"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-09-1143"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuroscience.2011.03.009"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-7843.2011.00843.x"},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.20994"},{"key":"e_1_2_7_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-7843.2011.00806.x"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1677\/joe.1.06433"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-7843.2010.00639.x"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(83)90303-4"},{"key":"e_1_2_7_35_1","doi-asserted-by":"publisher","DOI":"10.1124\/pr.110.003285"},{"key":"e_1_2_7_36_1","doi-asserted-by":"publisher","DOI":"10.1155\/2012\/473712"},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-2952(03)00548-3"},{"key":"e_1_2_7_38_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1025699932270"},{"key":"e_1_2_7_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-2952(00)00573-6"},{"key":"e_1_2_7_40_1","first-page":"1887","article-title":"Adenosine\u2010mediated killing of cultured epithelial cancer cells","volume":"60","author":"Barry CP","year":"2000","journal-title":"Cancer Res"},{"key":"e_1_2_7_41_1","first-page":"2602","article-title":"The extracellular fluid of solid carcinomas contains immunosuppressive concentrations of adenosine","volume":"57","author":"Blay J","year":"1997","journal-title":"Cancer Res"},{"key":"e_1_2_7_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2010.02.004"},{"key":"e_1_2_7_43_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.03-0011fje"},{"key":"e_1_2_7_44_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11010-008-9877-3"},{"key":"e_1_2_7_45_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.8.4941"},{"key":"e_1_2_7_46_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M401023200"},{"key":"e_1_2_7_47_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.167.1.197"},{"key":"e_1_2_7_48_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1476-5381.2009.00432.x"},{"key":"e_1_2_7_49_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI119833"},{"key":"e_1_2_7_50_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-540-89615-9_1"},{"key":"e_1_2_7_51_1","doi-asserted-by":"publisher","DOI":"10.1177\/026119290203000208"},{"key":"e_1_2_7_52_1","doi-asserted-by":"crossref","first-page":"973","DOI":"10.18388\/abp.2003_3627","article-title":"Degradation of extracellular nucleotides and their analogs in HeLa and HUVEC cell cultures","volume":"50","author":"Gendaszewska\u2010Darmach E","year":"2003","journal-title":"Acta Biochim Pol"},{"key":"e_1_2_7_53_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.10431"},{"key":"e_1_2_7_54_1","doi-asserted-by":"publisher","DOI":"10.3892\/ijmm.2012.1203"},{"key":"e_1_2_7_55_1","first-page":"199","article-title":"Regulation of cellular ATP release","volume":"118","author":"Fitz JG","year":"2007","journal-title":"Trans Am Clin Climatol Assoc"},{"key":"e_1_2_7_56_1","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.3104re1"},{"key":"e_1_2_7_57_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2004.11.027"},{"key":"e_1_2_7_58_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijdevneu.2010.06.001"},{"key":"e_1_2_7_59_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10585-007-9080-z"},{"key":"e_1_2_7_60_1","doi-asserted-by":"publisher","DOI":"10.1159\/000082930"},{"key":"e_1_2_7_61_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.abb.2010.05.001"},{"key":"e_1_2_7_62_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11302-006-9008-0"},{"key":"e_1_2_7_63_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1216013110"},{"key":"e_1_2_7_64_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2133.2007.07821.x"}],"container-title":["Basic &amp; Clinical Pharmacology &amp; Toxicology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fbcpt.12280","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/bcpt.12280","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,24]],"date-time":"2023-09-24T00:59:44Z","timestamp":1695517184000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/bcpt.12280"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,7,19]]},"references-count":63,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2015,1]]}},"alternative-id":["10.1111\/bcpt.12280"],"URL":"https:\/\/doi.org\/10.1111\/bcpt.12280","archive":["Portico"],"relation":{},"ISSN":["1742-7835","1742-7843"],"issn-type":[{"value":"1742-7835","type":"print"},{"value":"1742-7843","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,7,19]]},"assertion":[{"value":"2014-01-01","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-05-26","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-07-19","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}