J Vet Sci.  2013 Jun;14(2):207-214. 10.4142/jvs.2013.14.2.207.

Radiation up-regulates the expression of VEGF in a canine oral melanoma cell line

Affiliations
  • 1Clinic for Internal Medicine and Infectious Diseases, Institute of Immunology, University of Veterinary Medicine, 1210 Vienna, Austria. Irene.flickinger@vetmeduni.ac.at
  • 2Department for Pathobiology, University of Veterinary Medicine, 1210 Vienna, Austria.
  • 3Small Animal Clinic Hollabrunn, 2020 Hollabrunn, Austria.
  • 4Department for Natural Sciences, University of Veterinary Medicine, 1210 Vienna, Austria.

Abstract

To evaluate radiosensitivity and the effects of radiation on the expression of vascular endothelial growth factor (VEGF) and VEGF receptors in the canine oral melanoma cell line, TLM 1, cells were irradiated with doses of 0, 2, 4, 6, 8 and 10 Gray (Gy). Survival rates were then determined by a MTT assay, while vascular endothelial growth factor receptor (VEGFR)-1 and -2 expression was measured by flow cytometry and apoptotic cell death rates were investigated using an Annexin assay. Additionally, a commercially available canine VEGF ELISA kit was used to measure VEGF. Radiosensitivity was detected in TLM 1 cells, and mitotic and apoptotic cell death was found to occur in a radiation dose dependent manner. VEGF was secreted constitutively and significant up-regulation was observed in the 8 and 10 Gy irradiated cells. In addition, a minor portion of TLM 1 cells expressed vascular endothelial growth factor receptor (VEGFR)-1 intracellularly. VEGFR-2 was detected in the cytoplasm and was down-regulated following radiation with increasing dosages. In TLM 1 cells, apoptosis plays an important role in radiation induced cell death. It has also been suggested that the significantly higher VEGF production in the 8 and 10 Gy group could lead to tumour resistance.

Keyword

cell line; radiation; TLM 1; vascular endothelial growth factor

MeSH Terms

Animals
Apoptosis/*radiation effects
Cell Line, Tumor/radiation effects
Dogs
Dose-Response Relationship, Radiation
Enzyme-Linked Immunosorbent Assay/veterinary
Melanoma/genetics/metabolism
Mouth Neoplasms/genetics/metabolism
Radiation Tolerance
Tetrazolium Salts/metabolism
Thiazoles/metabolism
Up-Regulation/*radiation effects
Vascular Endothelial Growth Factor A/genetics/metabolism/*radiation effects
Vascular Endothelial Growth Factor Receptor-1/genetics/metabolism/*radiation effects
Vascular Endothelial Growth Factor Receptor-2/genetics/metabolism/*radiation effects
Tetrazolium Salts
Thiazoles
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factor Receptor-1
Vascular Endothelial Growth Factor Receptor-2
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