A simple method to induce hypoxia-induced vascular endothelial growth factor-A (VEGF-A) expression in T24 human bladder cancer cells

• Published in: In vitro cellular & developmental biology. Animal 2017-03, Vol.53 (3), p.272-276
• Main Authors: Cesário, Jonas Magno Santos, Brito, Rodrigo Barbosa Oliveira, Malta, Camila Soares, Silva, Chrisna Souza, Matos, Yves Silva Teles, Kunz, Tânia Cristina Macedo, Urbano, Jessica Julioti, Oliveira, Luis Vicente Franco, Dalboni, Maria Aparecida, Dellê, Humberto
• Format: Article
• Language: English
• Subjects: Animal Genetics and Genomics; Biomedical and Life Sciences; Cell Biology; Cell Culture; Cell Hypoxia - genetics; Cell Line, Tumor; CELLULAR PATHOLOGY/VIROLOGY; Developmental Biology; Gene Expression Regulation, Neoplastic; Humans; Hypoxia-Inducible Factor 1, alpha Subunit - biosynthesis; Hypoxia-Inducible Factor 1, alpha Subunit - genetics; Life Sciences; Neovascularization, Pathologic - genetics; Neovascularization, Pathologic - pathology; Oxygen - metabolism; RNA, Messenger - biosynthesis; Signal Transduction; Stem Cells; Urinary Bladder Neoplasms - genetics; Urinary Bladder Neoplasms - pathology; Vascular Endothelial Growth Factor A - biosynthesis; Vascular Endothelial Growth Factor A - genetics
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1007/s11626-016-0103-4

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 realtime 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.