Regulatory Regions required for β-Defensin-2 Up-regulation by Interleukin-1α in the Human Middle Ear Epithelial Cell Line
BACKGROUND AND OBJECTIVES: In the middle ear, the innate immunity is important because of less immune cell. Among the innate immunities beta-defensin-2 is known to play an important role in the immune function of middle ear. But, it is unclear about signal transduction pathway and gene regulatory region of human betadefensin-2 (hBD-2). MATERIALS AND METHOD: The expression of hBD-2 by IL-1a in HMEEC (Human Middle Ear Epithelial Cell) was detected by RT-PCR. The luciferase-expressing vector containing diverse lengths of the hBD-2 5' flanking region made by the progressive unidirectional deletion was transferred to HMEEC. We analyzed the function of 5’ flanking region by luciferase activity measured using luminometer after corresponding substrate was supplemented to the cell lysate. RESULTS: hBD-2 was upregulated by IL-1a in HMEEC-1. The treatment of IL-1a upregulates the activity of promoter 7.60±1.45 (average ± standard deviation) folded in 2.7kpb sized 5’ flanking region, 3.81 ± 0.78 folded in 1.1kbp, and 4.00 ± 0.73 folded in 500bp, respectively. CONCLUSION: These results indicate there are two effective gene regions that regulate hBD-2 expression by IL-1a between 2.7kbp and 1.1kbp, and 500bp upstream of the translation starting point of hBD-2 in HMEEC-1.
베타 디펜신-2, 유전자 조절 영역, 인터루킨-1, 중이, 면역계, Beta-defensin-2, Gene regulatory region, Interleukin-1, Middle ear, Immunity
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