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First published on June 13, 2005; DOI: 10.1124/mol.105.014357

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Received for publication May 2, 2005.
Revised June 10, 2005.
Accepted for publication June 13, 2005.

Increased Bcl2 expression by antisense oligoribonucleotides targeting the ARE motif

Laura Ghisolfi 1, Laura Papucci 2, Annamaria Bevilacqua 1, Gianfranco Canti 1, Giuseppe Tataranni 1, Andrea Lapucci 2, Nicola Schiavone 2, Sergio Capaccioli 2, Angelo Nicolin 1*

1 Pharmacology Department, University of Milan 2 Department of Experimental Pathology and Oncology, University of Florence

* Address correspondence to: E-mail: angelo.nicolin{at}unimi.it

Abstract

RNA has become a promising target for pharmacological purposes. Most current strategies are directed towards down-regulating its functions. Here we provide evidence of the up-regulation of messenger RNA in a sequence-specific manner. The b-ARE (adenine-uridine rich element) in the 3' untranslated region of the b-RNA (bcl2 RNA) that regulates the rate of RNA degradation has been targeted with three chemically modified oligoribonucleotides designed in the antisense orientation (asORNs). The three asORNs were studied by a cell-free degradation assay. All three slowed the rate of RNA decay in a dose-response fashion, they were specific to the b-ARE and two of them were individually effective. AsORNs were then transfected into the malignant cells in culture and b-RNA half-life was measured by real-time RT-PCR. We showed that by stabilizing b-RNA the three asORNs increased the expression of b-RNA and of the relevant protein in a dose-response fashion.


Key words: Antisense, Overexpression, RNA/siRNA, Oncogenes


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