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Published on: March 24, 2017
[An experimental study of inhibiting effect of decoy oligodeoxynucleotides on the gene expression of collagen in
Tian-zhi Hao1, Huan-ping Liang, Feng-lin Lv
1Department of neurosurgery, Research Institute of surgery, Daping Hospital, Third military Medical University, Chongqing 400042, PR China.
Objective:
To investigate the effect of activator protein-1 (AP-1) decoy-oligodeoxynucleotides (Decoy-ODNs) on the expression of fibroblast alpha2 type I collagen, so as to explore the gene therapy of pathologic scar.
Methods:
Decoy-ODNs targeting AP-1 were designed and synthesized. NIH3T3 cells were transfected by cationic liposomes. The distribution of Decoy-ODNs in the cells was investigated. The inhibiting effects of Decoy-ODNs on AP-1 were determined by electrophoretic mobility shift assay (EMSA). And the effects of Decoy-ODNs on the collagen synthesis in the cells were analyzed by RT-PCR.
Results:
AP-1 Decoy-ODNs could competitively inhibit the AP-1 in vitro activity. Cationic liposomes could play roles by effectively transfecting Decoy-ODNs into the plasma and nucleus. The mRNA expression of fibroblast alpha2 type I collagen decreased evidently after 24 hours of Decoy-ODNs action.
Conclusion:
Decoy-ODNs could inhibit the mRNA expression of fibroblast alpha2 type I collagen by antagonizing AP-1.
Insights
Activator protein-1 (AP-1) decoy-oligodeoxynucleotides (Decoy-ODNs) effectively inhibit fibroblast collagen production. This finding offers a potential gene therapy approach for managing pathologic scars.
Area of Science:
- Molecular Biology
- Gene Therapy
- Cell Biology
Context:
- Pathologic scars result in excessive collagen deposition.
- Fibroblast collagen synthesis is a key factor in scar formation.
- Activator protein-1 (AP-1) is a transcription factor involved in collagen gene expression.
Purpose:
- To investigate the efficacy of AP-1 decoy-oligodeoxynucleotides (Decoy-ODNs) in inhibiting fibroblast alpha2 type I collagen expression.
- To explore the potential of Decoy-ODNs as a gene therapy strategy for pathologic scars.
Summary:
- AP-1 Decoy-ODNs were synthesized and delivered into NIH3T3 cells using cationic liposomes.
- Electrophoretic mobility shift assay (EMSA) confirmed the inhibitory effect of Decoy-ODNs on AP-1 activity.
- RT-PCR analysis demonstrated a significant decrease in fibroblast alpha2 type I collagen mRNA expression after 24 hours of Decoy-ODN treatment.
Impact:
- Decoy-ODNs can effectively antagonize AP-1, leading to reduced collagen synthesis.
- This study provides a foundation for developing novel gene therapies targeting fibrotic conditions like pathologic scars.
- The findings highlight the therapeutic potential of modulating transcription factor activity in scar management.
