Complex host cell responses to antisense suppression of ACHE gene expression.
Antisense Nucleic Acid Drug Dev
; 11(1): 51-7, 2001 Feb.
Article
en En
| MEDLINE
| ID: mdl-11258621
3'-End-capped, 20-mer antisense oligodeoxynucleotides (AS-ODN) protected with 2'-O-methyl (Me) or phosphorothioate (PS) substitutions were targeted to acetylcholinesterase (AChE) mRNA and studied in PC12 cells. Me-modified AS-ODN suppressed AChE activity up to 50% at concentrations of 0.02-100 nM. PS-ODN was effective at 1-100 nM. Both AS-ODN displayed progressively decreased efficacy above 10 nM. In situ hybridization and confocal microscopy demonstrated dose-dependent decreases, then increases, in AChE mRNA. Moreover, labeling at nuclear foci suggested facilitated transcription or stabilization of AChE mRNA or both under AS-ODN. Intracellular concentrations of biotinylated oligonucleotide equaled those of target mRNA at extracellular concentrations of 0.02 nM yet increased only 6-fold at 1 microM ODN. Above 50 nM, sequence-independent swelling of cellular, but not nuclear, volume was observed. Our findings demonstrate suppressed AChE expression using extremely low concentrations of AS-ODN and attribute reduced efficacy at higher concentrations to complex host cell feedback responses.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Acetilcolinesterasa
/
Oligodesoxirribonucleótidos Antisentido
Límite:
Animals
Idioma:
En
Revista:
Antisense Nucleic Acid Drug Dev
Asunto de la revista:
BIOLOGIA MOLECULAR
/
BIOQUIMICA
/
FARMACOLOGIA
Año:
2001
Tipo del documento:
Article
País de afiliación:
Israel
Pais de publicación:
Estados Unidos