Combined p19Arf and interferon-beta gene transfer enhances cell death of B16 melanoma in vitro and in vivo

Author(s)

Merkel, C. A. ^{[1, 2, 3]} ; Medrano, R. F. V. ^[1] ; Barauna, V. G. ^[2] ; Strauss, B. E. ^{[1, 2, 3]}

Affiliation

^[1] Univ Sao Paulo, Sch Med, Inst Heart, Viral Vector Lab, BR-05403900 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Sch Med, Inst Heart, Lab Genet & Mol Cardiol LIM13, BR-05403900 Sao Paulo - Brazil

^[3] Univ Sao Paulo, Sch Med, Anim Care Facil, BR-05403900 Sao Paulo - Brazil

• Document type: Journal article
• Source: Cancer Gene Therapy; v. 20, n. 5, p. 317-325, MAY 2013.
• Web of Science Citations: 12

Abstract

Approximately 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. (AU)

• FAPESP's process: 10/03958-2 - Employment of IFN-beta and the p53/Arf pathway for the development of a new immunotherapy for melanoma
• Grantee: Ruan Felipe Vieira Medrano
• Support Opportunities: Scholarships in Brazil - Master