Corroborating these findings, Cunha-Filho et al (2010) and

Corroborating these findings, Cunha-Filho et al. (2010) and

Sciani et al. (2012) did not find hemolytic activity in amphibian skin secretions from R. crucifer, R. marina, R. schneideri and R. major at a concentration of 50 μg/mL, though secretions of R. jimi, R. margaritifer and Phyllomedusa hypochondrialis showed membrane disruption after 1 h incubation. Divergent results were seen with R. guttatus venom extracts, whereas all exhibited find more hemolytic potentiality, a contradictory finding when compared to that described by Sciani et al. (2012), who reported no membrane damage. It is likely that this difference should be correlated with range of concentrations used. The antiproliferative effects of the extracts were investigated on the basis of the incorporation of BrdU, a thymidine analog, into DNA, which occurs during the S phase of the cell cycle. R. marina extracts caused inhibition of DNA synthesis in HL-60 leukemia as evidenced by the decrease in BrdU incorporation, corroborating outcomes achieved with MTT and Alamar Blue™ CAL-101 ic50 assays. In fact, investigations have demonstrated that some toad skin secretions possess compounds able to induce cell cycle

arrest in G2/M phase, decrease cell viability, activate initiator and effector caspases and provoke morphological alterations (chromatin condensation, nuclear fragmentation, cytoplasm retraction, cell detachment, membrane blebs and apoptotic Rucaparib bodies) in prostate and breast carcinomas ( Yeh et al., 2003 and Sciani et al., 2012). Since cardiotonic steroids of two chemical classes, cardenolides (ouabain, for example) and bufadienolides,

bind specifically to the subunits of the sodium/potassium pump (Na+/K+-ATPase) ( Newman et al., 2008 and Gao et al., 2011), it is possible that the stimulation of apoptosis by bufadienolides is associated with this bioactivity. In summary, nine extracts of R. marina and R. guttatus venoms showed pronounced lethal and discriminating effects in tumor lines, especially those from R. marina, highlighting toad parotoid gland secretions as a promising source of novel lead anticancer compounds. HPLC and LC–MS analysis of the extracts of R. marina and R. guttatus venom showed significant differences between them, where four bufadienolides (1, 2, 3, and 4) were identified in different extracts from R. marina and only one (2) in R. guttatus. We are grateful to the Brazilian agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Mato Grosso (FAPEMAT), Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP) and Fundação de Amparo à Pesquisa do Estado do Piauí (FAPEPI) for financial support. The authors are indebted to Prof. Dr. M. L. dos Santos and Dr. G. A.

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