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Sequence specific antisense oligonucleotide analog interference with spermidine/spermine N1-acetyltransferase gene
M Fogel-Petrovic1, S Vujcic, R Häner
1Grace Cancer Drug Center, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
Anticancer Research
|September 1, 1996
Summary
Antisense oligonucleotide analogs (AOs) targeting the stop codon region of spermidine/spermine N1-acetyltransferase (SSAT) mRNA significantly suppressed SSAT induction by the polyamine analog DENSPM. However, this suppression did not fully inhibit DENSPM-induced cell growth inhibition.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Polyamines are crucial for cell growth, and their analogs, like N1,N11-diethylnorspermine (DENSPM), exhibit antiproliferative effects.
- DENSPM's antiproliferative action is linked to the induction of spermidine/spermine N1-acetyltransferase (SSAT), a complex gene response involving transcription, mRNA, and protein stabilization.
Purpose of the Study:
- To investigate the efficacy of SSAT-directed antisense oligonucleotide analogs (AOs) in preventing SSAT enzyme induction by DENSPM.
- To identify specific regions of SSAT mRNA targeted by AOs for effective suppression of DENSPM-induced enzyme activity.
Main Methods:
- Synthesis and evaluation of nine 18-mer fully phosphorothioate modified AOs targeting various regions of human SSAT mRNA.
- Treatment of MALME-3M human melanoma cells with AOs and DENSPM (10 microM for 6 hours).
- Quantification of SSAT activity, enzyme protein levels, and mRNA expression to assess AO efficacy.
Main Results:
- AOs targeting the stop codon region of SSAT mRNA were most effective in suppressing DENSPM-induced SSAT.
- AO-82 demonstrated a 70-80% suppression of SSAT activity, protein, and mRNA, suggesting interference with mRNA stabilization.
- Suppression of SSAT induction by AOs did not significantly alter the antiproliferative effects of DENSPM on cell growth.
Conclusions:
- Antisense oligonucleotide analogs targeting the SSAT mRNA stop codon region can effectively inhibit DENSPM-induced SSAT.
- The observed suppression of SSAT induction by AOs may not be sufficient to completely abrogate the antiproliferative effects of DENSPM.
- Further research may be needed to explore the precise role of SSAT induction in DENSPM's antiproliferative activity or to develop more potent AO strategies.