In vivo inhibition of tumor angiogenesis by a soluble VEGFR-2 fragment.
The interaction of vessel endothelial cell growth factor (VEGF) and its receptors (flt-1, FLK-1/KDR) regulates tumor angiogenesis. Therefore, blocking the binding of VEGF and the corresponding receptor has become critical for antitumor angiogenesis biological therapy. Our study extracted sFLK-1 fragment from embryo mouse liver using RT-PCR, recombined it to retrovirus vector, and transfected it to tumor cell lines (S180 and B16) by the liposome mediated method, then we observed the biological behavior of transgenic cells in vivo. The results are: (1) Fragment (1034 bp) was extracted from E9, E11 embryo mouse liver tissue, which was identified by sequence analysis. (2) This fragment was cloned to retrovirus vector (PLXSN vector), which was further transfected to tumor cells lines (S180 and B16). SDS-PAGE indicated the suspension of transgenic cells present sVEGFR-2(sFLK-1) fragment; Western blot identified it. (3) In vivo study showed that the weight and size of tumor in the group of transgenic cells were smaller than in control groups. Microvessel density (MVD) and FLK-1 expression were obviously different between transgenic and control groups, but there were no differences in VEGF expression between transgenic and control groups. In short, the isolated soluble VEGFR2 fragment transfected to tumor cells can be secreted to extracellular suspension and can inhibit tumor angiogenesis in vivo.
Kou
Baijun
B
Department of Pathology, College of Basic Medicine, Jilin University, Changchun 130021, China. koubaijun@yahoo.com
Li
Yulin
Y
Zhang
Lihong
L
Zhu
Guibin
G
Wang
Xinrui
X
Li
Yilei
Y
Xia
Jianxin
J
Shi
Yingai
Y
D000818:Animals; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D005354:Fibrosarcoma / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism; D008545:Melanoma / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009374:Neoplasms, Experimental / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D009389:Neovascularization, Pathologic / Q000378:metabolism* / Q000503:physiopathology; D010446:Peptide Fragments / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000502:physiology*
D010446:Peptide Fragments; D040301:Vascular Endothelial Growth Factor Receptor-2