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Ji Eun E Lee, Hye Shin Jeon, Jin Young Kim, Jae Ho Jung, Dong Hoon Shin, Min Kyu Shin; Induction of vascular endothelial growth factor and vascular endothelial growth factor receptor after repeated bevicizumab treatment in human umbilical vein endothelial cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3452.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in hypoxic human umbilical vein endothelial cells (HUVECs) after repeated anti-VEGF treatment.
HUVECs were incubated under the hypoxic environment with single or repeated treatment of bevacizumab at 1.0 or 2.5 mg/ml concentration. Treatment with identical volume of excipient was served as control. Expression of VEGF and phosphorylated VEGFR2 were assessed by enzyme-linked immuno-sorbant assay (ELISA) and western blot. Cytotoxicity and cell proliferation were also evaluated by MTT assay and immunocytochemistry of Ki-67.
VEGF secretion was significantly higher after the single treatment than control and the repeated treatment groups (P < 0.05). VEGFR2 expression increased significantly after both a single and repeated bevacizumab treatments than control (P < 0.05). VEGFR2 expression was higher after repeated treatment than a single treatment, and in lower concentration of bevacizumab than higher concentration (P < 0.05). Cytotoxicity or inhibition of proliferation by bevacizumab was not observed.
A single treatment of anti-VEGF induced secretion of VEGF from HUVECs, whereas repeated treatments increased expression of VEGFR2. Elevated receptor expression may increase responsiveness to VEGF, and would be one of the mechanisms for losing efficacy of anti-VEGF treatment in vivo.
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