Diana Matias1,2, Joana Balça-Silva1,3,4, Luiz Gustavo Dubois1, Bruno Pontes2, Valéria Pereira Ferrer1, Luciane Rosário1, Anália do Carmo3,5, Juliana Echevarria-Lima6, Ana Bela Sarmento-Ribeiro3,4,5, Maria Celeste Lopes3,7, Vivaldo Moura-Neto8. 1. Brain's Biomedicine Laboratory, Paulo Niemeyer State Brain Institute, Secretaria de Estado de Saúde do Rio de Janeiro, Rua do Resende 156, Rio de Janeiro, 20231-092, Rio de Janeiro, Brazil. 2. Institute of Biomedical Sciences at Federal University of Rio de Janeiro (ICB/UFRJ), Rio de Janeiro, 21941-902, Brazil. 3. Center for Neuroscience and Cell Biology and Institute for Biomedical Imaging and Life Sciences (CNC.IBILI), Rua Larga Faculdade de Medicina, Pólo I, 1° andar, 3004-504, Coimbra, Portugal. 4. Faculty of Medicine at University of Coimbra (FMUC), Pólo III - Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-354, Coimbra, Portugal. 5. Hospital Center and University of Coimbra (CHUC), Praceta Prof. Mota Pinto, 3000-075, Coimbra, Portugal. 6. Paulo de Góes Institute of Microbiology, Federal University of Rio de Janeiro, Rio de Janeiro, 21941-902, Brazil. 7. Faculty of Pharmacy at University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548, Coimbra, Portugal. 8. Brain's Biomedicine Laboratory, Paulo Niemeyer State Brain Institute, Secretaria de Estado de Saúde do Rio de Janeiro, Rua do Resende 156, Rio de Janeiro, 20231-092, Rio de Janeiro, Brazil. vivaldomouraneto@gmail.com.
Abstract
PURPOSE: Glioblastomas (GBM) comprise 17% of all primary brain tumors. These tumors are extremely aggressive due to their infiltrative capacity and chemoresistance, with glial-to-mesenchymal transition (GMT) proteins playing a prominent role in tumor invasion. One compound that has recently been used to reduce the expression of these proteins is shikonin (SHK), a naphthoquinone with anti-tumor properties. Temozolomide (TMZ), the most commonly used chemotherapeutic agent in GBM treatment, has so far not been studied in combination with SHK. Here, we investigated the combined effects of these two drugs on the proliferation and motility of GBM-derived cells. METHODS: The cytotoxic and proliferative effects of SHK and TMZ on human GBM-derived cells were tested using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), Ki67 staining and BrdU incorporation assays. The migration capacities of these cells were evaluated using a scratch wound assay. The expression levels of β3 integrin, metalloproteinases (MMPs) and GMT-associated proteins were determined by Western blotting and immunocytochemistry. RESULTS: We found that GBM-derived cells treated with a combination of SHK and TMZ showed decreases in their proliferation and migration capacities. These decreases were followed by the suppression of GMT through a reduction of β3 integrin, MMP-2, MMP-9, Slug and vimentin expression via inactivation of PI3K/AKT signaling. CONCLUSION: From our results we conclude that dual treatment with SHK and TMZ may constitute a powerful new tool for GBM treatment by reducing therapy resistance and tumor recurrence.
PURPOSE:Glioblastomas (GBM) comprise 17% of all primary brain tumors. These tumors are extremely aggressive due to their infiltrative capacity and chemoresistance, with glial-to-mesenchymal transition (GMT) proteins playing a prominent role in tumor invasion. One compound that has recently been used to reduce the expression of these proteins is shikonin (SHK), a naphthoquinone with anti-tumor properties. Temozolomide (TMZ), the most commonly used chemotherapeutic agent in GBM treatment, has so far not been studied in combination with SHK. Here, we investigated the combined effects of these two drugs on the proliferation and motility of GBM-derived cells. METHODS: The cytotoxic and proliferative effects of SHK and TMZ on human GBM-derived cells were tested using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), Ki67 staining and BrdU incorporation assays. The migration capacities of these cells were evaluated using a scratch wound assay. The expression levels of β3 integrin, metalloproteinases (MMPs) and GMT-associated proteins were determined by Western blotting and immunocytochemistry. RESULTS: We found that GBM-derived cells treated with a combination of SHK and TMZ showed decreases in their proliferation and migration capacities. These decreases were followed by the suppression of GMT through a reduction of β3 integrin, MMP-2, MMP-9, Slug and vimentin expression via inactivation of PI3K/AKT signaling. CONCLUSION: From our results we conclude that dual treatment with SHK and TMZ may constitute a powerful new tool for GBM treatment by reducing therapy resistance and tumor recurrence.
Authors: Suzana Assad Kahn; Deborah Biasoli; Celina Garcia; Luiz Henrique M Geraldo; Bruno Pontes; Morgana Sobrinho; Ana Carina Bon Frauches; Luciana Romão; Rossana C Soletti; Fernando dos Santos Assunção; Fernanda Tovar-Moll; Jorge Marcondes de Souza; Flavia R S Lima; Gregor Anderluh; Vivaldo Moura-Neto Journal: Curr Top Med Chem Date: 2012 Impact factor: 3.295
Authors: Roger Stupp; Monika E Hegi; Warren P Mason; Martin J van den Bent; Martin J B Taphoorn; Robert C Janzer; Samuel K Ludwin; Anouk Allgeier; Barbara Fisher; Karl Belanger; Peter Hau; Alba A Brandes; Johanna Gijtenbeek; Christine Marosi; Charles J Vecht; Karima Mokhtari; Pieter Wesseling; Salvador Villa; Elizabeth Eisenhauer; Thierry Gorlia; Michael Weller; Denis Lacombe; J Gregory Cairncross; René-Olivier Mirimanoff Journal: Lancet Oncol Date: 2009-03-09 Impact factor: 41.316
Authors: Frank B Furnari; Tim Fenton; Robert M Bachoo; Akitake Mukasa; Jayne M Stommel; Alexander Stegh; William C Hahn; Keith L Ligon; David N Louis; Cameron Brennan; Lynda Chin; Ronald A DePinho; Webster K Cavenee Journal: Genes Dev Date: 2007-11-01 Impact factor: 11.361
Authors: Luciana F Romão; Fabio A Mendes; Natalia M Feitosa; Jane Cristina O Faria; Juliana M Coelho-Aguiar; Jorge Marcondes de Souza; Vivaldo Moura Neto; José Garcia Abreu Journal: PLoS One Date: 2013-01-31 Impact factor: 3.240
Authors: Emma Camacho-Urkaray; Jorge Santos-Juanes; Francisco Borja Gutiérrez-Corres; Beatriz García; Luis M Quirós; Isabel Guerra-Merino; José Javier Aguirre; Iván Fernández-Vega Journal: Cell Oncol (Dordr) Date: 2017-12-07 Impact factor: 6.730