Effects of SVMPs Jararhagin and Berythractivase in Melanoma Cells: citotoxicity and cell cycle phases

Maria,DA.1; Ventura JS.1; Lapetina DL.1; Chudzinski-Tavassi AM.1

1 Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, Brasil.

Jararhagin is a 52 kDa metalloprotease from Bothropsjararaca snake venom, is a hemorrhagic and fibrino(geno)lytic toxin able to modulate the binding of von Willebrand factor A1 domain inhibiting platellet  aggregation induced by collagen.

Berythractivase is a 78 kDa metalloprotease from Bothrops erythromellas snake venom, is a prothrombin activator that does not promote hemorrhagic activity.

In this study we evaluated the anti-prolitevative action of both proteins on melanoma tumor cells regarding to citotoxic effects and cell cycle phases.

Confluent cultured of B16F10 melanoma cells were treated with different concentrations of Jararhagin (0.001-1,2uM) or Berythractivase (0.001 –0,6uM).The cytological alterations were analysed by inverse microscopy. Trypan blue test were used to evaluate cell viability and inhibition of adhesion was analysed by crystal violet and absorbance at 620nm. Phases of cell cycle were performed by flow cytometric on a Scalibur-BD (Becton-Dickson), using a Cell-Quest and Modi-fit Softwares.

The results showed that Jararhagin cause detachment by a dose and time dependent manner and this effect is reversible because the viability and grows capacity of the culture were maintained. On the other hand in high concentration Jararhagin induce citotoxicity decreasing significantly the S cell cycle phase. In lower concentration Berythractivase promotes detachment and induce significant arrest in GO/G1 demonstrating its anti-proliferative effect.

All together our results suggest that Berythractivase can modulate proliferation and growing of melanoma tumor cells.

Supported by FAPESP

Correspondence to: jsventura@ig.com.br