Double-stranded mols. 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 mols. recognizing Sp1 and NF-kB 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-kB binding sites inhibit the binding of NF-kB p52 and p50 transcription factors to target DNA mols. In this article we detd. the resistance of double-stranded decoy mols. based on PNA-DNA chimeras to exonucleases (both 3'®5' and 5'®3' exonucleases), endonucleases, and 5'-phosphatases. In addn., we performed expts. aimed at detg. the resistance of these mols. in cellular exts. and serum. Finally, we used liposomes as protective agents in exptl. conditions in which the decoy mols. employed were found to be unstable (high concns. of enzymes, cellular exts., or serum). The results obtained demonstrated that decoy mols. based on PNA-DNA chimeras are more resistant than DNA-based decoys to exo- and endonucleases, serum, and cellular exts. In addn., the resistance of DNA/PNA hybrids in the presence of high concns. of serum and cellular exts. 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 mols. 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
Saviano Michele;
2003-01-01
Abstract
Double-stranded mols. 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 mols. recognizing Sp1 and NF-kB 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-kB binding sites inhibit the binding of NF-kB p52 and p50 transcription factors to target DNA mols. In this article we detd. the resistance of double-stranded decoy mols. based on PNA-DNA chimeras to exonucleases (both 3'®5' and 5'®3' exonucleases), endonucleases, and 5'-phosphatases. In addn., we performed expts. aimed at detg. the resistance of these mols. in cellular exts. and serum. Finally, we used liposomes as protective agents in exptl. conditions in which the decoy mols. employed were found to be unstable (high concns. of enzymes, cellular exts., or serum). The results obtained demonstrated that decoy mols. based on PNA-DNA chimeras are more resistant than DNA-based decoys to exo- and endonucleases, serum, and cellular exts. In addn., the resistance of DNA/PNA hybrids in the presence of high concns. of serum and cellular exts. 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 mols. based on PNA-DNA chimeras for an efficient decoy treatment of tumor cells both in vitro and in vivo.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
