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《Chinese Pharmacological Bulletin》 2005-07
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Administration of a decoy against the CREB binding site suppressesthe expression of nNOS and fosB genes in morphine-dependent SK-N-SH cells

SU Yan-jun, CONG Bin, ZHANG Guo-zhong, ZHANG Jin, YAO Yu-xia, LI Shu-jin, FU Li-hong (Dept of Fornesic Science, Hebei Medical University,Shijiazhuang 050017,China)  
Aim To investigate the inhibitory effects of a synthetic CRE-transcription factor decoy oligodeoxynucleotide (CRE-decoy ODN) on the upregulation of the mRNA expression of nNOS and fosB in morphine-dependent SK-N-SH cells. Methods Morphine-dependent SK-N-SH cells were used. A synthetic single-stranded phosphorothioate oligodeoxynucleotide composed of the cAMP response element (CRE) sequence used as the CRE-decoy ODN in the presence of cationic lipid N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methylsulfate (DOTAP) was added to the culture medium. The effects of the CRE-decoy ODN on the DNA-binding activity of CREB, the expression of neuronal nitric oxide synthase (nNOS) and fosB mRNA were detected by electrophoresis mobility shift assay (EMSA) and RT-PCR respectively. Results Morphine dependence significantly activated the DNA-binding activity of CREB and the expression of nNOS and fosB mRNA. The CRE-decoy ODN could penetrate into SK-N-SH cells, specifically and stably suppress these indexes. Conclusion The CRE-decoy ODN can downregulate the expression of nNOS and fosB mRNA through specifically suppressing the DNA-binding activity of CREB in morphine-dependent SK-N-SH cells.
【Fund】: 河北省医学适用技术跟踪资助项目(No2001-54)
【CateGory Index】: R96
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