Inhibition of Sp1 activity by a decoy PNA-DNA chimera prevents urokinase receptor expression and migration of breast cancer cells(511 views) Zannetti A, Del Vecchio S, Romanelli A, Scala S, Saviano M, Cali' G, Stoppelli MP, Pedone C, Salvatore M
Keywords: Breast Cancer, Peptide Nucleic Acid, Pna-Dna Chimera, Sp1 Transcription Factor, Urokinase Receptor, Complementary Dna, Liposome, Oligonucleotide, Transcription Factor Sp1, Amino Acid Sequence, Article, Cancer Cell, Cancer Inhibition, Cell Migration, Cell Motility, Covalent Bond, Dna Strand, Down Regulation, Genetic Transfection, Human, Human Cell, In Vitro Study, Inhibition Kinetics, Metastasis, Priority Journal, Protein Binding, Protein Expression, Protein Function, Receptor Upregulation, Breast Neoplasms, Cell Line, Tumor, Cell Movement, Female, Cell Surface,
Affiliations: *** IBB - CNR ***
Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale Delle Ricerche, Via S. Pansini 5, 80131 Naples, Italy
Istituto di Endocrinologia Ed Oncologia Sperimentale, Consiglio Nazionale Delle Ricerche, Naples, Italy
Istituto Internazionale di Genetica e Biofisica, Consiglio Nazionale Delle Ricerche, Naples, Italy
Dipartimento di Scienze Biomorfologiche e Funzionali, Universita' Federico II, Naples, Italy
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Dignam, J. D., Lebovitz, R. M., Roeder, R. G., Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei (1983) Nucleic Acids Res, 11, pp. 1475-1489
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Mann, M. J., Dzau, V. J., Therapeutic applications of transcription factor decoy oligonucleotides (2000) J Clin Invest, 106, pp. 1071-1075
Cho, Y. S., Kim, M. K., Cheadle, C., Neary, C., Park, Y. G., Becker, K. G., A genomic-scale view of the cAMP response element-enhancer decoy: A tumor target-based genetic tool (2002) Proc Natl Acad Sci USA, 99, pp. 15626-15631
Wang, L. H., Yang, X. Y., Zhang, X., Mihalic, K., Xiao, W., Farrar, W. L., The cis decoy against the estrogen response element suppresses breast cancer cells via target disrupting c-fos not mitogen-activated protein kinase activity (2003) Cancer Res, 63, pp. 2046-2051
Mann, M. J., Whittemore, A. D., Donaldson, M. C., Belkin, M., Conte, M. S., Polak, J. F., Ex-vivo gene therapy of human vascular bypass grafts with E2F decoy: The PREVENT single-centre, randomised, controlled trial (1999) Lancet, 354, pp. 1493-1498
Novak, E. M., Metzger, M., Chammas, R., Da Costa, M., Dantas, K., Manabe, C., Downregulation of TNF-alpha and VEGF expression by Sp1 decoy oligodeoxynucleotides in mouse melanoma tumor (2003) Gene Ther, 10, pp. 1992-1997
Matrisian, L. M., Sledge Jr., G. W., Mohla, S., Extracellular proteolysis and cancer: Meeting summary and future directions (2003) Cancer Res, 63, pp. 6105-6109
Coussens, L. M., Fingleton, B., Matrisian, L. M., Matrix metalloproteinase inhibitors and cancer: Trials and tribulations (2002) Science, 295, pp. 2387-2392
Inhibition of Sp1 activity by a decoy PNA-DNA chimera prevents urokinase receptor expression and migration of breast cancer cells
Sp1 regulates the activation of many genes involved in tumor growth, apoptosis, and angiogenesis. We have previously shown the involvement of Sp1 in the up-regulation of urokinase receptor (uPAR) expression, a key molecule in tumor invasion and metastasis. Here, we investigated whether a marked down-regulation of Sp1 activity may inhibit uPAR expression and migration ability of MDA-MB-231 breast cancer cells. To this end, we tested the decoy ability of a novel peptide nucleic acid (PNA)-DNA chimera which carries a central DNA strand, containing Sp1-binding sequence, covalently linked to two PNA fragments at both ends (PNA-DNA-PNA, PDP). The chimera was synthesized, annealed with complementary DNA (PDP-DNA), and then tested for its ability to bind Sp1 both in vitro and in living MDA-MB-231 breast cancer cells in the presence of urokinase (uPA). This PDP-DNA decoy molecule efficiently competes for the binding to endogenous Sp1 in nuclear extracts, and upon transfection with liposomal vectors, causes a marked decrease of available Sp1 in both untreated and uPA-treated MDA-MB-231 cells. Accordingly, both uPA-dependent enhancement of uPAR expression and cell migration were strongly reduced in transfected cells. Interestingly, a detectable inhibitory effect is also observed in breast cancer cells exposed to PDP-DNA in the absence of transfection reagents. Finally, the inhibitory effect of PDP-DNA appeared to be stronger than that observed with oligonucleotides carrying Sp1 consensus sequence. Our findings show that this novel PNA-DNA chimera, containing Sp1 consensus sequence, effectively inhibits Sp1 activity, uPAR expression, and motility of breast cancer cells indicating its potential therapeutic use to prevent tumor dissemination. (c) 2005 Elsevier Inc. All rights reserved.
Inhibition of Sp1 activity by a decoy PNA-DNA chimera prevents urokinase receptor expression and migration of breast cancer cells
Cusanno F, Cisbani E, Colilli S, Fratoni R, Garibaldi F, Giuliani F, Gricia M, Lucentini M, Magliozzi ML, Santarivenere F, Torrioli S, Cinti MN, Pani R, Pellegrini R, Simonetti G, Schillaci O, Del Vecchio S, Salvatore M, Majewski S, De Vincentis G, Scopinaro F * Results of clinical trials with SPEM(278 views) Nucl Instrum Methods Phys Res Sect A, 2007 Feb 1; 497(1): 46-50. Impact Factor:3.221 ViewExport to BibTeXExport to EndNote