ABSTRACT
<p><b>OBJECTIVE</b>To verify whether the extracellular domain of kinase domain region (KDR) has anti-angiogenesis activity in vivo.</p><p><b>METHODS</b>cDNA was cloned into adeno-associated virus (AAV) vector pSNAV and transfected to baby hamster kidney (BHK) cells. Recombinant AAV was obtained from the cell culture supernatant after adding helper virus. Recombinant AAV-infected human bladder cancer EJ cell line (EJ cells) were injected subcutaneously into Balb-c nude mice. Tumor specimens were removed from the mice, paraffin-embedded and sliced, then stained by immunohistochemistry. Microvessel density (MVD) was determined under a microscope.</p><p><b>RESULTS</b>The tumor volume developed by EJ cells transfected with the extracellular domain of KDR was significantly smaller (1.70 +/- 0.18 cm(3)) compared with that in the control (5.62 +/- 0.67 cm(3)) (P < 0.05), although tumor developed to be detectable on almost the same time (14.7 +/- 2.4 days vs 14.1 +/- 3.2 days). Further, MVD in the experimental group was lower than that in the control (41.3 +/- 4.8 vs 6.2 +/- 2.1, P < 0.05).</p><p><b>CONCLUSION</b>The extracellular domain of KDR could be expressed in nude mouse bladder cancer tissue and inhibit tumor angiogenesis.</p>
Subject(s)
Animals , Cricetinae , Female , Mice , Cloning, Molecular , Dependovirus , Genetics , Endothelial Growth Factors , Metabolism , Genetic Therapy , Intercellular Signaling Peptides and Proteins , Metabolism , Lymphokines , Metabolism , Mice, Inbred BALB C , Mice, Nude , Neovascularization, Pathologic , Urinary Bladder Neoplasms , Therapeutics , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factor Receptor-2 , Genetics , Vascular Endothelial Growth FactorsABSTRACT
Objective To verify whether the extracellular domain of flt 1 has anti angionesis activity in vivo. Methods A recombinant plasmid pcDNA 3.1/flt 1 n3 was transfected into the human bladder carcinoma EJ cell by lipofectamin, which was constructed by inserting the N terminal first three IgG like domain of VEGF receptor flt 1 (flt 1 n3 ) into eukaryotic expression vector pcDNA 3.1. Results The rflt 1 was functionally expressed in stably transfected EJ and the product secreted in the medium could be specifically bind to rhVEGF165.The result showed that the bladder cancer transfected with rflt 1 had a lower microvessle density than the control. Conclusions It is proved that the expressed rflt 1 n3 can inhibit tumor growth and angiogenesis in nude mice model.