Limits...
Antiproliferative effect of the jararhagin toxin on B16F10 murine melanoma.

Maria DA, da Silva MG, Correia Junior MC, Ruiz IR - BMC Complement Altern Med (2014)

Bottom Line: Malignant melanoma is a less common but highly dangerous form of skin cancer; it starts in the melanocytes cells found in the outer layer of the skin.Proliferative rate was assessed by staining with 5,6-carboxyfluoresceindiacetate succinimidyl ester, showing a significant decrease in proliferation at all concentrations of both toxins.In vivo treatment of the toxins was observed reduction in the incidence of nodules, and metastasis and antiproliferative inhibition capacity.

View Article: PubMed Central - PubMed

Affiliation: Biochemistry and Biophysics Laboratory, Butantan Institute, Av, Vital Brasil 1500, CEP 05503-900 Sao Paulo, SP, Brazil. durvanei.maria@butantan.gov.br.

ABSTRACT

Background: Malignant melanoma is a less common but highly dangerous form of skin cancer; it starts in the melanocytes cells found in the outer layer of the skin. Jararhagin toxin, a metalloproteinase isolated from Bothrops jararaca snake venom acts upon several biological processes, as inflammation, pain, platelet aggregation, proliferation and apoptosis, though not yet approved for use, may one day be employed to treat tumors.

Methods: B16F10 murine melanoma cells were treated with jararhagin (jara), a disintegrin-like metalloproteinase isolated from Bothrops jararaca snake venom, and jari (catalytic domain inactivated with 1,10-phenanthroline). Viability and adhesion cells were evaluated by MTT assay. The expression of caspase-3 active, phases of the cell cycle and apoptosis were assessed by flow cytometry. We analyze in vivo the effects of jararhagin on melanoma growth, apoptosis and metastasis.

Results: The tumor cells acquired round shapes, lost cytoplasmic expansions, formed clusters in suspension and decreased viability. Jari was almost 20 times more potent toxin than jara based on IC50 values and on morphological changes of the cells, also observed by scanning electron microscopy. Flow cytometry analysis showed 48.3% decrease in the proliferation rate of cells and 47.2% increase in apoptosis (jara) and necrosis (jari), following 1.2 μM jara and 0.1 μM jari treatments. Caspase-3 activity was increased whereas G0/G1 cell cycle phase was on the decline. Proliferative rate was assessed by staining with 5,6-carboxyfluoresceindiacetate succinimidyl ester, showing a significant decrease in proliferation at all concentrations of both toxins.

Conclusions: In vivo treatment of the toxins was observed reduction in the incidence of nodules, and metastasis and antiproliferative inhibition capacity. This data strengthens the potential use jararhagin as an anti-neoplastic drug.

Show MeSH

Related in: MedlinePlus

Cell cycle analysis of B16F10 cells treated with jara and jari. Cells were stained with iodide propidium for DNA content analysis by flow cytometry. The bars represent the proportions of G2/M proliferative cells; in phase S synthesis; G0/G1 quiescent cell, and debris in sub-G1. The figure shows a significant decrease in the percentage of cells in G0/G1 phase and a subsequent increase in sub-G1 phase with increased concentration of jara and jari. Data represents mean ± SD from three independent experiments. *Significantly different from control *p < 0.05; **p < 0.01 and ***p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4289281&req=5

Fig7: Cell cycle analysis of B16F10 cells treated with jara and jari. Cells were stained with iodide propidium for DNA content analysis by flow cytometry. The bars represent the proportions of G2/M proliferative cells; in phase S synthesis; G0/G1 quiescent cell, and debris in sub-G1. The figure shows a significant decrease in the percentage of cells in G0/G1 phase and a subsequent increase in sub-G1 phase with increased concentration of jara and jari. Data represents mean ± SD from three independent experiments. *Significantly different from control *p < 0.05; **p < 0.01 and ***p < 0.001.

Mentions: The distribution of populations in the cell cycle phases was checked after jara and jari treatments. The percentage of cells in G0/G1 phases was significantly reduced by both toxins; the percentage of cells in S phase was significantly decreased only with 0.4 μM jari treatment. No significant alterations on the distribution of G2/M cells were induced by both toxins. Sub-G1 cell populations (debris and fragmented DNA) were significantly increased after jara and especially jari treatments (Figure 7).Figure 7


Antiproliferative effect of the jararhagin toxin on B16F10 murine melanoma.

Maria DA, da Silva MG, Correia Junior MC, Ruiz IR - BMC Complement Altern Med (2014)

Cell cycle analysis of B16F10 cells treated with jara and jari. Cells were stained with iodide propidium for DNA content analysis by flow cytometry. The bars represent the proportions of G2/M proliferative cells; in phase S synthesis; G0/G1 quiescent cell, and debris in sub-G1. The figure shows a significant decrease in the percentage of cells in G0/G1 phase and a subsequent increase in sub-G1 phase with increased concentration of jara and jari. Data represents mean ± SD from three independent experiments. *Significantly different from control *p < 0.05; **p < 0.01 and ***p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4289281&req=5

Fig7: Cell cycle analysis of B16F10 cells treated with jara and jari. Cells were stained with iodide propidium for DNA content analysis by flow cytometry. The bars represent the proportions of G2/M proliferative cells; in phase S synthesis; G0/G1 quiescent cell, and debris in sub-G1. The figure shows a significant decrease in the percentage of cells in G0/G1 phase and a subsequent increase in sub-G1 phase with increased concentration of jara and jari. Data represents mean ± SD from three independent experiments. *Significantly different from control *p < 0.05; **p < 0.01 and ***p < 0.001.
Mentions: The distribution of populations in the cell cycle phases was checked after jara and jari treatments. The percentage of cells in G0/G1 phases was significantly reduced by both toxins; the percentage of cells in S phase was significantly decreased only with 0.4 μM jari treatment. No significant alterations on the distribution of G2/M cells were induced by both toxins. Sub-G1 cell populations (debris and fragmented DNA) were significantly increased after jara and especially jari treatments (Figure 7).Figure 7

Bottom Line: Malignant melanoma is a less common but highly dangerous form of skin cancer; it starts in the melanocytes cells found in the outer layer of the skin.Proliferative rate was assessed by staining with 5,6-carboxyfluoresceindiacetate succinimidyl ester, showing a significant decrease in proliferation at all concentrations of both toxins.In vivo treatment of the toxins was observed reduction in the incidence of nodules, and metastasis and antiproliferative inhibition capacity.

View Article: PubMed Central - PubMed

Affiliation: Biochemistry and Biophysics Laboratory, Butantan Institute, Av, Vital Brasil 1500, CEP 05503-900 Sao Paulo, SP, Brazil. durvanei.maria@butantan.gov.br.

ABSTRACT

Background: Malignant melanoma is a less common but highly dangerous form of skin cancer; it starts in the melanocytes cells found in the outer layer of the skin. Jararhagin toxin, a metalloproteinase isolated from Bothrops jararaca snake venom acts upon several biological processes, as inflammation, pain, platelet aggregation, proliferation and apoptosis, though not yet approved for use, may one day be employed to treat tumors.

Methods: B16F10 murine melanoma cells were treated with jararhagin (jara), a disintegrin-like metalloproteinase isolated from Bothrops jararaca snake venom, and jari (catalytic domain inactivated with 1,10-phenanthroline). Viability and adhesion cells were evaluated by MTT assay. The expression of caspase-3 active, phases of the cell cycle and apoptosis were assessed by flow cytometry. We analyze in vivo the effects of jararhagin on melanoma growth, apoptosis and metastasis.

Results: The tumor cells acquired round shapes, lost cytoplasmic expansions, formed clusters in suspension and decreased viability. Jari was almost 20 times more potent toxin than jara based on IC50 values and on morphological changes of the cells, also observed by scanning electron microscopy. Flow cytometry analysis showed 48.3% decrease in the proliferation rate of cells and 47.2% increase in apoptosis (jara) and necrosis (jari), following 1.2 μM jara and 0.1 μM jari treatments. Caspase-3 activity was increased whereas G0/G1 cell cycle phase was on the decline. Proliferative rate was assessed by staining with 5,6-carboxyfluoresceindiacetate succinimidyl ester, showing a significant decrease in proliferation at all concentrations of both toxins.

Conclusions: In vivo treatment of the toxins was observed reduction in the incidence of nodules, and metastasis and antiproliferative inhibition capacity. This data strengthens the potential use jararhagin as an anti-neoplastic drug.

Show MeSH
Related in: MedlinePlus