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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: CANCER CELL INTERNATIONAL; v. 20, n. 1 DEC 17 2020.
• Citações Web of Science: 0

Resumo

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)

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