Resistance of decoy PNA-DNA chimeras to enzymatic degradation in cellular extracts and serum(372 views) Borgatti M, Romanelli A, Saviano M, Pedone C, Lampronti I, Breda L, Nastruzzi C, Bianchi N, Mischiati C, Gambari R
Oncology Research (ISSN: 0965-0407), 2003; 13(5): 279-287.
Keywords: Decoy, Peptide Nucleic Acid (pna), Peptide Nucleic Acid (pna)-Dna Chimeras, Cell Extract, Endonuclease, Exonuclease, Immunoglobulin Enhancer Binding Protein, Liposome, Protein P50, Transcription Factor, Transcription Factor Decoy, Transcription Factor Sp1, Unclassified Drug, Exodeoxyribonuclease, Oligonucleotide, Phosphatase, Article, Binding Site, Cancer Invasion, Controlled Study, Dna Binding, Dna Flanking Region, Enzyme Activity, Enzyme Degradation, Gene Targeting, Human, Human Cell, In Vitro Study, In Vivo Study, Priority Journal, Protein Localization, Tumor Growth, Animal, Blood, Cattle, Cell Line, Chemistry, Dose Response, Enzyme Activation, Metabolism, Physiology, Dose-Response Relationship, Phosphoric Monoester Hydrolases,
Affiliations: *** IBB - CNR ***
Dept. of Biochem. and Molec. Biology, Ferrara University, Ferrara, Italy
Inst. of Biostructure and Bioimaging, CNR, Napoli, Italy
Dept. Pharmaceut. Chem. and Technol., University of Perugia, Perugia, Italy
Biotechnology Center, Ferrara University, Ferrara, Italy
Dipto. di Biochim. e Biol. Molec., Via L. Borsari n.46, 44100 Ferrara, Italy
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Cho-Chung, Y. S., Park, Y. G., Nesterova, M., Lee, Y. N., Cho, Y. S., CRE-decoy oligonucleotide-inhibition of gene expression and tumor growth (2000) Mol. Cell. Biochem., 212, pp. 29-34
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Kozlov, I. A., Kubareva, E. A., Ivanovskaya, M. G., Shabarova, Z. A., Design of new reagents on the base of DNA duplexes for irreversible inhibition of transcription factor NF-kappa B (1997) Antisense Nucleic Acid Drug Dev., 7, pp. 279-289
Lim, C. S., Jabrane-Ferrat, N., Fontes, J. D., Okamoto, H., Garovoy, M. R., Peterlin, B. M., Hunt, C. A., Sequence-independent inhibition of RNA transcription by DNA dumbbells and other decoys (1997) Nucleic Acids Res., 25, pp. 575-580
Romano, M. F., Lamberti, A., Bisogni, R., Tassone, P., Pagnini, D., Storti, G., Del Vecchio, L., Venuta, S., Enhancement of cytosine arabinoside-induced apoptosis in human myeloblastic leukemia cells by NF-kappa B/Rel-specific decoy oligodeoxynucleotides (2000) Gene Ther., 7, pp. 1234-1237
Sharma, H. W., Narayanan, R., The NF-kappaB transcription factor in oncogenesis (1996) Anticancer Res., 16, pp. 589-596
Sharma, H. W., Perez, J. R., Higgins-Sochaski, K., Hsiao, R., Narayanan, R., Transcription factor decoy approach to decipher the role of NF-kappa B in oncogenesis (1996) Anticancer Res., 16, pp. 61-69
Cho-Chung, Y. S., Nesterova, M., Becker, K. G., Srivastava, R., Park, Y. G., Lee, Y. N., Cho, Y. S., Cheadle, C., Dissecting the circuitry of protein kinase A and cAMP signaling in cancer genesis: Antisense, microarray, gene overexpression, and transcription factor decoy (2002) Ann. NY Acad. Sci., 968, pp. 22-36
Nielsen, P. E., Egholm, M., Berg, R. H., Buchardt, O., Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide (1991) Science, 254, pp. 1497-1500
Demidov, V. V., Potaman, V. N., Frank-Kamenetskii, M. D., Egholm, M., Buchard, O., Sonnichsen, S. H., Nielsen, P. E., Stability of peptide nucleic acids in human serum and cellular extracts (1994) Biochem. Pharmacol., 48, pp. 1310-1313
Scarf, S., Giovine, M., Gasparini, A., Damonte, G., Millo, E., Pozzolini, M., Benatti, U., Modified peptide nucleic acids are internalized in mouse macrophages RAW 264. 7 and inhibit inducible nitric oxide synthase (1999) FEBS Lett., 451, pp. 264-268
Resistance of decoy PNA-DNA chimeras to enzymatic degradation in cellular extracts and serum
Double-stranded molecules based on peptide nucleic acids (PNAs)-DNA chimeras carrying binding sites for known transcription factors could be of great interest in decoy pharmacotherapy of neoplastic diseases. For instance, decoy molecules recognizing Sp1 and NF-kappaB transcription factors were found to inhibit tumor cell growth and invasion activity. In this respect, we have recently found that double-stranded PNA-DNA chimeras carrying NF-kappaB binding sites inhibit the binding of NF-kappaB p52 and p50 transcription factors to target DNA molecules. In this article we determined the resistance of double-stranded decoy molecules based on PNA-DNA chimeras to exonucleases (both 3'-->5' and 5'-->3' exonucleases), endonucleases, and 5'-phosphatases. In addition, we performed experiments aimed at determining the resistance of these molecules in cellular extracts and serum. Finally, we used liposomes as protective agents in experimental conditions in which the decoy molecules employed were found to be unstable (high concentrations of enzymes, cellular extracts, or serum). The results obtained demonstrated that decoy molecules based on PNA-DNA chimeras are more resistant than DNA-based decoys to exo- and endonucleases, serum, and cellular extracts. In addition, the resistance of DNA/PNA hybrids in the presence of high concentrations of serum and cellular extracts was increased after complexation to cationic liposomes, due to the fact that double-stranded PNA-DNA-PNA chimeras bind to these delivery systems. The results obtained in the present study support the proposal of molecules based on PNA-DNA chimeras for an efficient decoy treatment of tumor cells both in vitro and in vivo.
Resistance of decoy PNA-DNA chimeras to enzymatic degradation in cellular extracts and serum
No results.
Resistance of decoy PNA-DNA chimeras to enzymatic degradation in cellular extracts and serum