A high-efficacy antisense RIalpha poly-DNP 21-nt RNA.
The antisense inhibitor poly-2'-O-(2,4-dinitrophenyl)-5'-GGCUGCGUGCCUCCUCACUGG (antisense poly-DNP RNA-21) has been synthesized by in vitro transcription followed by chemical derivatization. Its base sequence is complementary to that of nucleotides 110-130 in the mRNA of the regulatory RIalpha subunit of PKA (RIalpha/PKA), which is overexpressed in MCF-7 breast cancer cells and A549 lung cancer cells. The bioavailable and RNase-resistant antisense poly-DNP RNA-21 was found to inhibit cell growth with 50% inhibitory concentration (IC50) values of 0.05 nM in MCF-7 cells and 4 nM in A549 cells. The control 21-nt RNAs with the same poly-DNP oligonucleotide (ODN) platform but with scrambled, sense, or mismatched base sequence are inactive. Treatment of MCF-7 cells with antisense poly-DNP RNA-21 abolishes both the steady-state concentration of RIalpha mRNA and the synthesis of RIalpha protein. At sufficiently high concentration, antisense poly-DNP RNA-21 selectively kills the targeted cancer cells by inducing apoptosis. The observed sequence specificity and extremely low IC50 values of antisense poly-DNP RNA-21 suggest that it is a promising candidate for in vivo testing as an effective anticancer agent.
Shen
Long
L
Bioenergetics Laboratory, Natural Sciences Complex, University at Buffalo, SUNY, Buffalo, NY 14260-3000, USA.
Chen
Xiaolan
X
Wang
Jui H
JH
D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics*; D006131:Growth Inhibitors / Q000494:pharmacology*; D006801:Humans; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit; D006131:Growth Inhibitors; D020320:Oligoribonucleotides, Antisense; C517834:PRKAR1A protein, human; D017868:Cyclic AMP-Dependent Protein Kinases