Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na+/K+-ATPase beta 2: potential molecular entities to treat invasive brain cancer

Author(s)

Barreto, Natalia ^{[1, 2]} ; Caballero, Marcus ^{[1, 2]} ; Bonfanti, Amanda Pires ^{[1, 2]} ; Pinheiro de Mato, Felipe Cezar ^{[1, 2]} ; Munhoz, Jaqueline ^{[1, 2]} ; da Rocha-e-Silva, Thomaz A. A. ^[3] ; Sutti, Rafael ^[4] ; Vitorino-Araujo, Joao Luiz ^[5] ; Verinaud, Liana ^[2] ; Raposo, Catarina ^[1]

Affiliation

^[1] Univ Estadual Campinas UNICAMP, Fac Ciencias Farmaceut, BR-13083865 Campinas, SP - Brazil

^[2] Univ Estadual Campinas, Inst Biol, Dept Biol Estrut & Func, Sao Paulo - Brazil

^[3] Fac Israelita Ciencias Saude Albert Einstein, Sao Paulo, SP - Brazil

^[4] Santa Casa Sao Paulo, Fac Ciencias Med, Sao Paulo, SP - Brazil

^[5] Fac Ciencias Med Santa Casa Sao Paulo, Disciplina Neurocirurgia, Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: CANCER CELL INTERNATIONAL; v. 20, n. 1 DEC 17 2020.
• Web of Science Citations: 0

Abstract

BackgroundGlioblastoma (GB) cells have the ability to migrate and infiltrate the normal parenchyma, leading to the formation of recurrent tumors often adjacent to the surgical extraction site. We recently showed that Phoneutria nigriventer spider venom (PnV) has anticancer effects mainly on the migration of human GB cell lines (NG97 and U-251). The present work aimed to investigate the effects of isolated components from the venom on migration, invasiveness, morphology and adhesion of GB cells, also evaluating RhoA-ROCK signaling and Na+/K+-ATPase beta 2 (AMOG) involvement.MethodsHuman (NG97) GB cells were treated with twelve subfractions (SFs-obtained by HPLC from PnV). Migration and invasion were evaluated by scratch wound healing and transwell assays, respectively. Cell morphology and actin cytoskeleton were shown by GFAP and phalloidin labeling. The assay with fibronectin coated well plate was made to evaluate cell adhesion. Western blotting demonstrated ROCK and AMOG levels and a ROCK inhibitor was used to verify the involvement of this pathway. Values were analyzed by the GraphPad Prism software package and the level of significance was determinate using one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparisons test.ResultsTwo (SF1 and SF11) of twelve SFs, decreased migration and invasion compared to untreated control cells. Both SFs also altered actin cytoskeleton, changed cell morphology and reduced adhesion. SF1 and SF11 increased ROCK expression and the inhibition of this protein abolished the effects of both subfractions on migration, morphology and adhesion (but not on invasion). SF11 also increased Na+/K+-ATPase beta 2.ConclusionAll components of the venom were evaluated and two SFs were able to impair human glioblastoma cells. The RhoA effector, ROCK, was shown to be involved in the mechanisms of both PnV components. It is possible that AMOG mediates the effect of SF11 on the invasion. Further investigations to isolate and biochemically characterize the molecules are underway. (AU)

• FAPESP's process: 15/04194-0 - Identification of new molecules with chemotherapeutic effect in human glioma and characterization of the mechanism
• Grantee: Catarina Raposo Dias Carneiro
• Support type: Research Grants - Young Investigators Grants
• FAPESP's process: 15/06134-4 - Multi-User Equipment approved in grant 2014/03002-7: cell imaging multi-mode reader
• Grantee: Marcelo Bispo de Jesus
• Support type: Multi-user Equipment Program
• FAPESP's process: 17/05402-0 - Identification and characterization of spider Phoneutria nigriventer isolated venom, with chemotherapeutic effect on brain tumor cells
• Grantee: Amanda Pires Bonfanti
• Support type: Scholarships in Brazil - Technical Training Program - Technical Training
• FAPESP's process: 17/16196-2 - Analysis of the effects and mechanisms of the Phoneutria nigriventer spider venom on morphology and migration of tumor cells
• Grantee: Natália Barreto dos Santos
• Support type: Scholarships in Brazil - Master
• FAPESP's process: 17/24331-7 - Analysis of the effect and mechanism of action of P. nigriventer venom and its isolated toxins on the development of brain tumor implanted in mice
• Grantee: Marcus Vinícius Campoy Chichizola Caballero Alves
• Support type: Scholarships in Brazil - Master
• FAPESP's process: 19/10003-3 - Molecular characterization of human gliomas and identification of neoplasms responsible for new biopharmaceuticals obtained from animal venom: a translational approach
• Grantee: Natália Barreto dos Santos
• Support type: Scholarships in Brazil - Doctorate
• FAPESP's process: 18/23559-7 - Cell therapy with macrophages and NK cells modulated ex vivo by peptide isolated from animal venom: a new approach in immunotherapy for cancer
• Grantee: Amanda Pires Bonfanti
• Support type: Scholarships in Brazil - Doctorate