title: A simple method to induce hypoxia-induced vascular endothelial growth factor-A (VEGF-A) expression in T24 human bladder cancer cells.
abstract: Angiogenesis is an essential process for the establishment, development, and dissemination of several malignant tumors including bladder cancer. The hypoxic condition promotes the stabilization of hypoxia-inducible factor 1 alpha (HIF-1α), which translocates to the nucleus to mediate angiogenic factors including the vascular endothelial growth factor A (VEGF-A). AnaeroGen system was developed for microbiology area to create a low oxygen tension required to the growth of anaerobic bacteria. Here, we hypothesized the use of AnaeroGen system to induce hypoxia in T24 human bladder carcinoma cells, in order to promote the overexpression of VEGF-A. T24 cells were cultured in six-well plates containing McCoy medium. Exposures of T24 cells to hypoxia for 1, 8, 24, and 48 h were performed using the Oxoid AnaeroGen system, while T24 cells under normoxia were used as control. The expression of VEGF-A and HIF-1α was analyzed by real-time PCR. ELISA for HIF-1α was carried out. The VEGF-A expression increased significantly by Oxoid AnaeroGen-induced hypoxia in a time-depending manner, reaching the peak in 48 h of hypoxia. Although HIF-1α mRNA was not changed, HIF-1α protein was increased in the presence of hypoxia, reaching a peak at 8 h. These results demonstrated that the Oxoid AnaeroGen system is a simple method to expose T24 cells to hypoxia and efficiently to upregulate VEGF expression in T24 cells.
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil.
authors.forename: Jonas Magno Santos
authors.initials: JM
authors.lastname: Cesário
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil.
authors.forename: Rodrigo Barbosa Oliveira
authors.initials: RB
authors.lastname: Brito
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil.
authors.forename: Camila Soares
authors.initials: CS
authors.lastname: Malta
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil.
authors.forename: Chrisna Souza
authors.initials: CS
authors.lastname: Silva
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil.
authors.forename: Yves Silva Teles
authors.initials: YS
authors.lastname: Matos
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil.
authors.forename: Tânia Cristina Macedo
authors.initials: TC
authors.lastname: Kunz
authors.affiliation: Sleep Laboratory, Rehabilitation Sciences Master Degree and PhD Program, Nove de Julho University (UNINOVE), São Paulo, Brazil.
authors.forename: Jessica Julioti
authors.initials: JJ
authors.lastname: Urbano
authors.affiliation: Sleep Laboratory, Rehabilitation Sciences Master Degree and PhD Program, Nove de Julho University (UNINOVE), São Paulo, Brazil.
authors.forename: Luis Vicente Franco
authors.initials: LV
authors.lastname: Oliveira
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil.
authors.forename: Maria Aparecida
authors.initials: MA
authors.lastname: Dalboni
authors.affiliation: Postgraduate Program in Medicine, Universidade Nove de Julho (UNINOVE), Rua Vergueiro, 235, 2° subsolo, São Paulo, São Paulo, 01504-001, Brazil. hdelle@usp.br.
authors.forename: Humberto
authors.initials: H
authors.lastname: Dellê
mesh_terms: D015687:Cell Hypoxia / Q000235:genetics*; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis* / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000473:pathology; D010100:Oxygen / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D015398:Signal Transduction; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D010100:Oxygen
keywords: Angiogenesis; Bladder carcinoma; HIF-1α; Hypoxia; Oxoid AnaeroGen; VEGF-A