Intercellular trafficking and enhanced in vivo antitumour activity of a non-virally delivered P27-VP22 fusion protein.
VP22, a structural protein from herpes simplex virus type I, exhibits the unique property of intercellular trafficking. This protein is exported from primary expressing cells and subsequently imported into neighbouring cells. This property is conserved when VP22 is genetically fused to a protein, making it a promising tool to enhance the delivery of a gene product. We chose to study the intercellular transport and biological effect of a fusion protein between the putative tumour suppressor gene p27(Kip1) and VP22. We show that in vitro, P27VP22 is able to spread as efficiently as VP22. Functionality of the P27VP22 protein was demonstrated by its ability to inhibit cyclin/CDK2 complexes activity. In proliferation and clonogenicity assays, transfection with the P27VP22 plasmid resulted in a stronger cell growth inhibition when compared to transfection with the p27(Kip1) vector. In vivo, sub cutaneous tumours established in nude mice were injected with naked DNA encoding P27 or P27VP22. Our results show that P27VP22 can spread in vivo and that injections of the P27VP22 plasmid resulted in a significantly greater antitumour activity than injections of the P27 plasmid. This study confirms the usefulness of VP22-mediated delivery and suggests that P27VP22 may have applications in cancer gene therapy.
Zavaglia
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Groupe de Recherche sur le Cancer du Pumon, Equipe INSERM 9924, Institut Albert Bonnoit, La Tronche cedex, France.
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Eymin
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Tenaud
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D000818:Animals; D039481:CD11b Antigen / Q000378:metabolism; D019556:COS Cells; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D018797:Cell Cycle Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D017874:Immediate-Early Proteins; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000628:therapy*; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D025521:Tumor Suppressor Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
D039481:CD11b Antigen; C493961:Cdkn1b protein, mouse; D018797:Cell Cycle Proteins; C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D011993:Recombinant Fusion Proteins; D025521:Tumor Suppressor Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; C078424:EUS1 protein, Equine herpesvirus 1; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases