| Summary: | 碩士 === 靜宜大學 === 化粧品科學系 === 104 === Elastic fibers are extracellular matrix component of the skin, which contributing to skin elasticity and tenacity, and in the process of formation of elastic fibers, the microfibrillar-associated protein 4 (MFAP4) in which is particularly important, such as the lack MFAPA4 will not form mature and compact structure of elastic fibers. Earlier study has revealed that MFAP4 can protect the skin from UVB-mediated photo-damage and photo-aging, which also prevent the degradation of extracellular matrix to enhance elastic fibers stability and skin elasticity. In this study, we use the secreted alkaline phosphatase (SEAP) and green fluorescent protein (GFP) reporter gene systems to analyze the predicted human MFAP4 gene promoter regions. Moreover, some compounds can control the expression of elastin and fibrillin-1 will also be used to investigate the effect on the regulation of MFAP4 gene. The analysis of MFAP4 promoter sequence found that it had the characteristic of TATA-less and the sequence containing Initiator element (Inr), Motif ten element (MTE) and Downstream promoter element (DPE). Analysis indicated that it is a core promoter. Moreover, within 250 base pair from the transcription start site, we also predicted some elements in the promoter region, including GC box, GATA box and CAAT box. The length of 0.5k, 1k and 2k base pair MFAP4 gene promoter in human fibroblasts cells (Detroit 551) can successful initiation of transcription and with promoter activity assay represents that 0.5k base pair of the promoter its transcriptional activity is relatively low. In addition, the transcriptional activities of 1k and 2k base pair promoter are not much difference. The MFAP4 gene promoters in mouse fibroblasts cells (NIH/3T3), human melanoma cells (MeWo) and mouse melanoma cells (B16-F10) are lacking of activity. Therefore, the results indicated that MFAP4 promoter is species-specific and cell type-specific one.
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