Antitumor potential of the myotoxin BthTX-I from Bothrops jararacussu snake venom: evaluation of cell cycle alterations and death mechanisms induced in tumor cell lines

Abstract

Abstract\ud \ud Background\ud Phospholipases A2 (PLA2s) are abundant components of snake venoms that have been extensively studied due to their pharmacological and pathophysiological effects on living organisms. This study aimed to assess the antitumor potential of BthTX-I, a basic myotoxic PLA2 isolated from Bothrops jararacussu venom, by evaluating in vitro processes of cytotoxicity, modulation of the cell cycle and induction of apoptosis in human (HL-60 and HepG2) and murine (PC-12 and B16F10) tumor cell lines.\ud \ud \ud Methods\ud The cytotoxic effects of BthTX-I were evaluated on the tumor cell lines HL-60 (promyelocytic leukemia), HepG2 (human hepatocellular carcinoma), PC-12 (murine pheochromocytoma) and B16F10 (murine melanoma) using the MTT method. Flow cytometry technique was used for the analysis of cell cycle alterations and death mechanisms (apoptosis and/or necrosis) induced in tumor cells after treatment with BthTX-I.\ud \ud \ud Results\ud It was observed that BthTX-I was cytotoxic to all evaluated tumor cell lines, reducing their viability in 40 to 50 %. The myotoxin showed modulating effects on the cell cycle of PC-12 and B16F10 cells, promoting delay in the G0/G1 phase. Additionally, flow cytometry analysis indicated cell death mainly by apoptosis. B16F10 was more susceptible to the effects of BthTX-I, with ~40 % of the cells analyzed in apoptosis, followed by HepG2 (~35 %), PC-12 (~25 %) and HL-60 (~4 %).\ud \ud \ud Conclusions\ud These results suggest that BthTX-I presents antitumor properties that may be useful for developing new therapeutic strategies against cancer.The authors would like to thank the financial support provided by the State\ud of São Paulo Research Foundation (FAPESP, grants n. 2010/03243-43 and\ud 2011/23236-4), the Coordination for the Improvement of Higher Education\ud Personnel (CAPES) and the National Council for Scientific and Technological\ud Development (CNPq process n. 476932/2012-2). We are also grateful to\ud Fabiana Rosseto Morais, from FCFRP-USP, for the technical assistance in the\ud flow cytometry analyses. Thanks are also due to the Center for the Study of\ud Venoms and Venomous Animals (CEVAP) of UNESP for enabling the publication\ud of this special collection (CNPq process 469660/2014-7)