Abstract
Immunohistochemical analysis has demonstrated that the human IFI16 gene, in addition to the hematopoietic tissues, is highly expressed in endothelial cells and squamous stratified epithelia. In this study, we have developed a reliable HSV-derived replication-defective vector (TO-IFI16) to efficiently transduce IFI16 into primary human umbilical vein endothelial cells (HUVEC), which are usually poorly transfectable. HUVEC infection with TO-IFI16 virus suppressed endothelial migration, invasion and formation of capillary-like structures in vitro. In parallel, sustained IFI16 expression inhibited HUVEC cell cycle progression, accompanied by significant induction of p53, p21, and hypophosphorylated pRb. Further support for the involvement of these pathways in IFI16 activity came from the finding that infection with TO-IFI16 virus does not impair the in vitro angiogenic activity and cell cycle progression of HUVEC immortalized by HPV16 E6/E7 oncogenes, which are known to inactivate both p53 and pRb systems. This use of a reliable viral system for gene delivery into primary human endothelial cells assigns a potent angiostatic activity to an IFN-inducible gene, namely IFI16, and thus throws further light on antiangiogenic therapy employing IFNs.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Angiogenesis Inhibitors / genetics*
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Angiogenesis Inhibitors / therapeutic use
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Capillaries / cytology
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Capillaries / growth & development*
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Capillaries / metabolism
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Cell Count
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Cell Cycle / genetics
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Cell Division / drug effects
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Cell Line, Transformed
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Cell Movement / genetics
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Cyclin-Dependent Kinase Inhibitor p21
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Cyclins / metabolism
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Endothelial Cells / cytology
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Endothelial Cells / metabolism*
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Endothelial Cells / virology
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Gene Expression Regulation, Neoplastic / genetics
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Genetic Vectors / therapeutic use*
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Humans
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Interferons / metabolism
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Interferons / pharmacology
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Neovascularization, Pathologic / genetics*
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Neovascularization, Pathologic / therapy
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Nuclear Proteins*
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Oncogene Proteins, Viral / genetics
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Papillomavirus E7 Proteins
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Phosphoproteins*
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Proteins / genetics*
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Proteins / therapeutic use
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Repressor Proteins*
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Retinoblastoma Protein / metabolism
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Simplexvirus / genetics
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Transduction, Genetic / methods*
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Tumor Suppressor Protein p53 / metabolism
Substances
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Angiogenesis Inhibitors
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CDKN1A protein, human
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Cyclin-Dependent Kinase Inhibitor p21
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Cyclins
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E6 protein, Human papillomavirus type 16
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Nuclear Proteins
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Oncogene Proteins, Viral
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Papillomavirus E7 Proteins
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Phosphoproteins
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Proteins
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Repressor Proteins
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Retinoblastoma Protein
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Tumor Suppressor Protein p53
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oncogene protein E7, Human papillomavirus type 16
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IFI16 protein, human
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Interferons