title: Potentiation of ribonuclease cytotoxicity by a poly(amidoamine) dendrimer.
abstract: Variants of bovine pancreatic ribonuclease (RNase A) engineered to evade the endogenous ribonuclease inhibitor protein (RI) are toxic to human cancer cells. Increasing the basicity of these variants facilitates their entry into the cytosol and thus increases their cytotoxicity. The installation of additional positive charge also has the deleterious consequence of decreasing ribonucleolytic activity or conformational stability. Here, we report that the same benefit can be availed by co-treating cells with a cationic dendrimer. We find that adding the generation 2 poly(amidoamine) dendrimer in trans increases the cytotoxicity of RI-evasive RNase A variants without decreasing their activity or stability. The increased cytotoxicity is not due to increased RI-evasion or cellular internalization, but likely results from improved translocation into the cytosol after endocytosis. These data indicate that co-treatment with highly cationic molecules could enhance the efficacy of ribonucleases as chemotherapeutic agents.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706, USA.
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Ellis
authors.forename: Megan L
authors.initials: ML
authors.lastname: Hornung
authors.forename: Ronald T
authors.initials: RT
authors.lastname: Raines
mesh_terms: D000818:Animals; D002412:Cations; D002417:Cattle; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D050091:Dendrimers / Q000737:chemistry* / Q000633:toxicity; D004359:Drug Therapy, Combination; D014644:Genetic Variation; D006801:Humans; D020128:Inhibitory Concentration 50; D020014:K562 Cells; D008958:Models, Molecular; D009369:Neoplasms / Q000188:drug therapy; D011073:Polyamines / Q000737:chemistry* / Q000633:toxicity; D012259:Ribonuclease, Pancreatic / Q000737:chemistry* / Q000235:genetics / Q000627:therapeutic use
chemical_list: D002412:Cations; D050091:Dendrimers; C531249:Poly(amidoamine); D011073:Polyamines; D012259:Ribonuclease, Pancreatic