title: Novel RasGRF1-derived Tat-fused peptides inhibiting Ras-dependent proliferation and migration in mouse and human cancer cells.
abstract: Mutations of RAS genes are critical events in the pathogenesis of different human tumors and Ras proteins represent a major clinical target for the development of specific inhibitors to use as anticancer agents. Here we present RasGRF1-derived peptides displaying both in vitro and in vivo Ras inhibitory properties. These peptides were designed on the basis of the down-sizing of dominant negative full-length RasGRF1 mutants. The over-expression of these peptides can revert the phenotype of K-RAS transformed mouse fibroblasts to wild type, as monitored by several independent biological readouts, including Ras-GTP intracellular levels, ERK activity, morphology, proliferative potential and anchorage independent growth. Fusion of the RasGRF1-derived peptides with the Tat protein transduction domain allows their uptake into mammalian cells. Chemically synthesized Tat-fused peptides, reduced to as small as 30 residues on the basis of structural constraints, retain Ras inhibitory activity. These small peptides interfere in vitro with the GEF catalyzed nucleotide dissociation and exchange on Ras, reduce cell proliferation of K-RAS transformed mouse fibroblasts, and strongly reduce Ras-dependent IGF-I-induced migration and invasion of human bladder cancer cells. These results support the use of RasGRF1-derived peptides as model compounds for the development of Ras inhibitory anticancer agents.
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milano, Italy. elena.sacco@unimib.it
authors.forename: Elena
authors.initials: E
authors.lastname: Sacco
authors.forename: David
authors.initials: D
authors.lastname: Metalli
authors.forename: Michela
authors.initials: M
authors.lastname: Spinelli
authors.forename: Romilde
authors.initials: R
authors.lastname: Manzoni
authors.forename: Maria
authors.initials: M
authors.lastname: Samalikova
authors.forename: Rita
authors.initials: R
authors.lastname: Grandori
authors.forename: Andrea
authors.initials: A
authors.lastname: Morrione
authors.forename: Silvio
authors.initials: S
authors.lastname: Traversa
authors.forename: Lilia
authors.initials: L
authors.lastname: Alberghina
authors.forename: Marco
authors.initials: M
authors.lastname: Vanoni
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D016503:Drug Delivery Systems; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D014162:Transfection; D018631:ras Proteins / Q000037:antagonists & inhibitors*; D020666:ras-GRF1 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D010446:Peptide Fragments; D010455:Peptides; D011993:Recombinant Fusion Proteins; D020666:ras-GRF1; D054322:tat Gene Products, Human Immunodeficiency Virus; C497137:tat peptide (47-57), Human immunodeficiency virus 1; D018631:ras Proteins