PMID: 16630405Apr 25, 2006Paper

Enhancement of antiangiogenic effects of human canstatin with a hypoxia-regulated transgene vector in lung cancer model

The Cancer Journal
Yuying LiGuo-Ming Wu

Abstract

Canstatin, a newly identified antiangiogenesis protein, has a potent inhibitory effect on the proliferation and growth of endothelial cells. To enhance the expression and antiangiogenic effects of canstatin in solid tumors, we constructed a eukaryotic expression vector that encodes human canstatin cDNA downstream from nine copies of the hypoxia-response element. Canstatin complementary DNA from adult liver tissues was cloned into the mammalian expression vector pCMV-Script. Nine copies of the hypoxia-response element were ligated upstream from the canstatin gene near the cytomegalovirus promoter. The recombinant vector, pCMV9Cans, was transformed into A549 cells by cationic liposomes. The transformed cells were cultured under oxic and anoxic conditions. We detected canstatin messenger ribonucleic acid and protein expression in transformed cells by TaqMan polymerase chain reaction and Western blot analysis, respectively. Human umbilical vein endothelial cells were cocultured with recombinant vector transformed A549 cells using Transwell plates under oxic and anoxic conditions. The proliferation and apoptosis of the cocultured endothelial cells were evaluated with 3H-thymidine incorporation and terminal deoxynucleotidyl-mediated ...Continue Reading

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