TY - JOUR
T1 - Woodchuck VEGF (wVEGF) characteristics
T2 - Model for angiogenesis and human hepatocellular carcinoma directed therapies
AU - Huang, Huayi
AU - Salavaggione, Oreste
AU - Rivera, Lee
AU - Mukherjee, Sarbajit
AU - Brekken, Rolf A
AU - Tennant, Bud
AU - Iyer, Renuka
AU - Adjei, Araba
N1 - Funding Information:
This study was supported by American Cancer Society MRSG-08-096-01-CCE , Roswell Park Alliance Grant (PI Iyer) and Roswell Park Research Institute Health Research Incorporated account 71–1020.
PY - 2019/1
Y1 - 2019/1
N2 - Vascular endothelial growth factor (VEGF) stimulates angiogenesis. Human hepatocellular carcinoma (HCC) is a VEGF-driven tumor often associated with chronic hepatitis B or C virus infection. The woodchuck is a well-characterized model of hepatitis B virus related HCC and a valuable tool for translational studies of novel VEGF targeted agents. We cloned the cDNA encoding woodchuck VEGF (wVEGF), transiently expressed it in COS cells and functionally characterized the recombinant protein. The open reading frame of wVEGF contained 645 nucleotides encoding a protein of 214 amino acids. Two protein bands (17 and 25 kDa) were detected in conditioned media of wVEGF expressing COS-1 cells and a single band of 25 kDa was identified in cell lysates. Addition of recombinant wVEGF to COS cells enhanced cell proliferation and stimulated VEGFR2, Akt, ERK1/2, and FAK phosphorylation. Sunitinib, a tyrosine kinase inhibitor, inhibited wVEGF- induced VEGFR2 phosphorylation in a dose-dependent manner. Finally, development of HCC in woodchucks was accompanied by increased laminin and PECAM1 expressing vessels, VEGFR2 expression, increased ligation of VEGF to VEGFR2, and a decrease in collagen IV-positive blood vessels. Our results suggest that woodchuck model can be used further to study angiogenesis and the effect of VEGF directed therapies in human HCC.
AB - Vascular endothelial growth factor (VEGF) stimulates angiogenesis. Human hepatocellular carcinoma (HCC) is a VEGF-driven tumor often associated with chronic hepatitis B or C virus infection. The woodchuck is a well-characterized model of hepatitis B virus related HCC and a valuable tool for translational studies of novel VEGF targeted agents. We cloned the cDNA encoding woodchuck VEGF (wVEGF), transiently expressed it in COS cells and functionally characterized the recombinant protein. The open reading frame of wVEGF contained 645 nucleotides encoding a protein of 214 amino acids. Two protein bands (17 and 25 kDa) were detected in conditioned media of wVEGF expressing COS-1 cells and a single band of 25 kDa was identified in cell lysates. Addition of recombinant wVEGF to COS cells enhanced cell proliferation and stimulated VEGFR2, Akt, ERK1/2, and FAK phosphorylation. Sunitinib, a tyrosine kinase inhibitor, inhibited wVEGF- induced VEGFR2 phosphorylation in a dose-dependent manner. Finally, development of HCC in woodchucks was accompanied by increased laminin and PECAM1 expressing vessels, VEGFR2 expression, increased ligation of VEGF to VEGFR2, and a decrease in collagen IV-positive blood vessels. Our results suggest that woodchuck model can be used further to study angiogenesis and the effect of VEGF directed therapies in human HCC.
KW - Hepatitis B virus
KW - Hepatocellular carcinoma (HCC)
KW - Vascular endothelial growth factor (VEGF)
KW - Vascular endothelial growth factor receptor 2 (VEGFR2)
KW - Woodchuck
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U2 - 10.1016/j.abb.2018.11.008
DO - 10.1016/j.abb.2018.11.008
M3 - Article
C2 - 30439360
AN - SCOPUS:85056754552
SN - 0003-9861
VL - 661
SP - 97
EP - 106
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
ER -