Increased Bcl2 expression by antisense oligoribonucleotides targeting the adenine-uridine-rich element motif.
RNA has become a promising target for pharmacological purposes. Most current strategies are directed toward down-regulating its functions. In this study, we provide evidence of the up-regulation of messenger RNA in a sequence-specific manner. The bcl2 (b)-ARE (adenine-uridine-rich element) in the 3'-untranslated region of the b-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 reverse transcriptase-polymerase chain reaction. 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.
Ghisolfi
Laura
L
Department of Pharmacology, University of Milan, Via Vanvitelli 32, 20129 Milan, Italy.
Papucci
Laura
L
Bevilacqua
Annamaria
A
Canti
Gianfranco
G
Tataranni
Giuseppe
G
Lapucci
Andrea
A
Schiavone
Nicola
N
Capaccioli
Sergio
S
Nicolin
Angelo
A
D000225:Adenine / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016372:RNA, Antisense / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D014529:Uridine / Q000378:metabolism*
D019253:Proto-Oncogene Proteins c-bcl-2; D016372:RNA, Antisense; D000225:Adenine; D014529:Uridine