Sistema FMUSP-HC: Faculdade de Medicina da Universidade de São Paulo (FMUSP) e Hospital das Clínicas da FMUSPMERKEL, C. A.MEDRANO, R. F. V.BARAUNA, V. G.STRAUSS, B. E.2013-09-232013-09-232013CANCER GENE THERAPY, v.20, n.5, p.317-325, 20130929-1903https://observatorio.fm.usp.br/handle/OPI/2462Approximately 90% of melanomas retain wild-type p53, a characteristic that may help shape the development of novel treatment strategies. Here, we employed an adenoviral vector where transgene expression is,controlled by p53 to deliver the p19 alternate reading frame (An) and interferon-beta (IFN beta) complementary DNAs in the B16 mouse model of melanoma. In vitro, cell death was enhanced by combined gene transfer (63.82 +/- 15.30% sub-GO cells); yet introduction of a single gene resulted in significantly fewer hypoploid cells (37.73 +/- 7.3% or 36.96 +/- 11.58%, p19Arf or IFN beta, respectively, P < 0.05). Annexin V staining and caspase-3 cleavage indicate a cell death mechanism consistent with apoptosis. Using reverse transcriptase quantitative PCR, we show that key transcriptional targets of p53 were upregulated in the presence of p19Arf, although treatment with IFN beta did not alter expression of the genes studied. In situ gene therapy revealed significant inhibition of subcutaneous tumors by IFN beta (571 +/- 25 mm(3)) or the combination of p19Arf and IFN beta (489 +/- 124 mm(3)) as compared with the LacZ control (1875 +/- 33 mm(3), P < 0.001); whereas p19Arf yielded an intermediate result (1053 +/- 169 mm(3), P < 0.01 vs control). However, only the combination was associated with increased cell death and prolonged survival (P < 0.01). As shown here, the combined transfer of p19Arf and IFN beta using p53-responsive vectors enhanced cell death both in vitro and in vivo.engrestrictedAccessp53p14Arf/p19Arfinterferonmelanomaadenoviruscell deathadvanced malignant-melanomaifn-betaadenoviral vectorantiviral defensetumor suppressionstabilizes p53phase-iitherapymurineexpressionarticleCopyright NATURE PUBLISHING GROUP10.1038/cgt.2013.23Biotechnology & Applied MicrobiologyOncologyGenetics & HeredityMedicine, Research & Experimental