Effect of Transcription Factor Decoy for NF-κB on the TNF-α Induced Cytokine and ICAM-1 Expression in Cultured HaCaT cells.
10.5021/ad.2003.15.3.93
- Author:
Kyu Suk LEE
;
Jee Ook KIM
;
Byung Chun KIM
;
Young Wook RYOO
- Publication Type:Original Article
- Keywords:
NF-κB decoy;
Cytokine;
In vitro
- MeSH:
Binding Sites;
Biological Therapy;
Cytokines;
DNA;
Electrophoretic Mobility Shift Assay;
Gene Expression;
Humans;
In Vitro Techniques;
Intercellular Adhesion Molecule-1*;
Interleukin-1;
Interleukin-6;
Liposomes;
Oligodeoxyribonucleotides;
Oligonucleotides;
Psoriasis;
RNA, Messenger;
Skin;
Skin Diseases;
Transcription Factors*;
Transfection;
Vascular Cell Adhesion Molecule-1
- From:Annals of Dermatology
2003;15(3):93-98
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: Psoriasis is the most prevalent T-cell-mediated inflammatory skin disease in humans. Numerous cytokines and adhesion molecules are expressed in the skin lesion of psoriasis such as TNF-α, IL-1, IL-6, VCAM-1 and ICAM-1. All of them contain at least one binding site for the transcription factor NF-κB. TNF-α activates NF-κB and many other transcription factors. Thus, transcription and expression of many genes involved in the inflammatory process may be influenced by TNF-α. OBJECTIVE: The purpose of this study was to study the effect of synthetic double-stranded DNA with high affinity for the NF-κB binding site on the TNF-α induced proinflammatory cytokines and ICAM-1 gene expression in the HaCaT cells. MATERIAL AND METHODS: We examined whether inhibition of NF-κB activity by oligodeoxynucleotides (ODN) decoy for NF-κB blocks TNF-α induced cytokines such as IL-la, IL-1 a, IL-6 and ICAM-1 expression with electrophoretic mobility shift assay (EMSA) and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In EMSA, TNF-α treatment (10 ng/ml) induced the activation of NF-κB. The NF-κB binding activity in the TNF-α treated HaCaT cells increased 5.0-fold compared to non-treated group. Next, we examined the effect of liposome mediated NF-κB decoy oligonucleotides (ODN) transfection. After transfection of the NF-κB decoy ODN, TNF-α increased NF-κB binding activity to 1.9-fold of non-treated group. Accordingly the transfection of NF-κB decoy ODN inhibited the TNF-α induced NF-κB binding activity up to 63%. RT-PCR analysis revealed that the transfection of NF-κB decoy ODN inhibited TNF-α induced cytokines and ICAM-1 mRNA expression. CONCLUSION: Taken together, our results suggest the potential utility of NF-κB decoy technique for biologic therapy of psoriasis.
