Summary
In order to explore the role of activator protein-1 (AP-1) in the transcription of interleukin-5 (IL-5) gene regulated by protein kinase C (PKC) signal in peripheral blood T lymphocytes from asthmatic patient, T lymphocytes were isolated and purified from peripheral blood of each asthmatic patient. The T lymphocytes were randomly divided into 4 groups: group A (blank control), group B (treated with PKC agonist phorbol 12-myristate 13-acetate (PMA)), Group C (treated with PMA and AP-1 cis-element decoy oligodeoxynucleotides (decoy ODNs)), and group D (treated with PMA and AP-1 mutant decoy ODNs). The ODNs were transfected into the T cells of group C and D by cation liposome respectively. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to assess IL-5 mRNA expression, and electrophoretic mobility shift assays (EMSA) for the activation of AP-1. The results showed that the activation of AP-1 (88 003. 58±1 626. 57) and the expression of IL-5 mRNA (0. 8300±0. 0294) in T lymphocytes stimulated with PMA were significantly higher than these in blank control (20 888. 47±1103. 56 and 0. 3050±0. 0208, respectively,P<0.01), while the indexes (23 219. 83±1 024. 86 and 0. 3425±0. 0171 respectively) of T lymphocytes stimulated with PMA and AP-1 decoy ODNs were significantly inhibited, as compared with group B (P<0.01). The indexes (87 107.41±1 342. 92 and 0. 8225±0 0222, respectively) in T lymphocytes stimulated with PMA and AP-1 mutant decoy ODNs did not exhibit significant changes, as compared with group B (P>0.05). The significant positive correlation was found between the activation of AP-1 and the expression of IL-5 mRNA (P<0.01). It was concluded that AP-1 might participate in the signal transduction of PKC-triggered transcription of IL-5 gene in asthmatic T lymphocytes. This suggests the activation of PKC/AP-1 signal transduction cascade of T lymphocytes may play an important role in the pathogenesis of asthma.
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References
O’Byrne P M, Inman M D, Parameswaran K. The trials and tribulations of IL-5, eosinophils, and allergic asthma. J Allergy Clin Immunol, 2001, 108:503
Mori A, Kaminuma O, Mikami Tet al. Transcriptional control of the IL-5 gene by human helper T cells: IL-5 synthesis is regulated independently from IL-2 or IL-4 synthesis. J Allergy Clin Immunol, 1999, 103: S429
Ogwa K, Kaminuma O, Okudaira Heet al. Transcriptional regulation of the IL-5 gene in peripheral T cells of asthmatic patients. Clin Exp Immunol, 2002, 130: 475
Liu C, Lu J, Tan Jet al. Human interleukin-5 expression is synergistically regulated by histone acetyltransferase CBP/p300 and transcription factors C/EBP, NFAT and AP-1. Cytokine, 2004, 27: 93
Krappmann D, Patke A, Heissmeyer Vet al. B-cell receptor- and phorbol ester-induced NF-kB and c-Jun N-Terminal kinase activation in B cells requires novel protein kinase C’s. Mol Cell Biol, 2001, 21: 6640
Xiong W, Xu Y, Zhang Zet al. Nuclear factor-kappaB in signal conduction of protein kinase C in T lymphocytes from an asthmatic guinea pig model. Chin Med J, 2002, 115: 685
Andrews N C, Faller D V. A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. Nucleic Acids Res, 1991, 19: 2499
Jans D A, Xiao C Y, Lam M H. Nuclear targeting signal recognition: a key control point in nuclear transport? Bioessays, 2000, 22: 532
Bielinska A, Shivdasani R A, Zhang Let al. Regulation of gene expression with double-stranded phosphorothioate oligonucleotides. Science, 1990, 250: 997
Morishita R, Gibbons G H, Horiuchi Met al. A gene therapy strategy using a transcription factor decoy of the E2F binding site inhibits smooth muscle proliferationin vivo. Proc Natl Acad Sci USA, 1995, 92: 5855
Ahn J D, Morishita R, Kaneda Yet al. Transcription factor decoy for AP-1 reduces mesangial cell proliferation and extracellular matrix production in vitro and vivo. Gene Ther, 2004, 11: 916
Karin M, Liu Z, Zandi E. AP-1 function and regulation. Curr Opin Cell Biol, 1997, 9: 240
Chinenov Y, Kerppola T K. Close encounters of many kinds: Fos-Jun interactionsthat mediate transcription regulatory specificity. Oncogene, 2001, 20: 2438
Dzau V J. Transcription factor decoy. Circ Res, 2002, 90: 1234
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Guo Qi, male, born in 1968, M. D., Ph. D.
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Qi, G., Yongjian, X. & Zhexiang, Z. Role of activator protein-1 in the transcription of interleukin-5 gene regulated by protein kinase C signal in asthmatic human T lymphocytes. Current Medical Science 25, 147–150 (2005). https://doi.org/10.1007/BF02873562
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DOI: https://doi.org/10.1007/BF02873562