順位因子Octamer參與脂多醣類刺激RAW264.7細胞resistin基因表現之探討

An octamer element is required for the regulation of LPS-induced resistin gene expression in RAW264.7 cells

陳嘉綾(Chia-Ling Chen)

指導教授 ： 呂紹俊

國立臺灣大學/醫學院/生物化學暨分子生物學研究所/碩士(2004年)

https://doi.org/10.6342/NTU.2004.10160

摘要

雖然最早的發現顯示肥胖可能是藉由resistin造成胰島素抗拒，但之後部分研究並不完全支持這個結果。實驗室之前以大鼠注射LPS(lipopolysaccharide)來模擬發炎反應，發現LPS會增加脂肪組織及白血球resistin mRNA的表現量。小鼠巨噬細胞RAW264.7細胞也會受LPS影響而使resistin mRNA明顯增加。由resistin mRNA在人類各組織表現量的分析發現，在bone marrow的resistin mRNA表現量遠大於其他組織。綜合這些結果顯示resistin可能參與發炎反應。因此本篇論文主要在探討RAW264.7細胞受LPS刺激引起resistin mRNA表現量增加的分子機制。
由實驗室之前的研究結果已知在RAW264.7細胞resistin promoter活性會受LPS刺激而增高，promoter上 -1037至-969及-968至-660區域對於LPS刺激的反應是必須的，因此將Resistin promoter(-1037/+61)-Luc質體做不同長度的internal deletion (

關鍵字

脂多醣類

並列摘要

From our previous studies, levels of resistin mRNA expression were significantly increased in both epididymal white adipose tissue and white blood cells after intraperitioneal injection of lipopolysaccharide (LPS). Furthermore, we found that resistin mRNA was expressed in human peripheral blood mononuclear cells and THP-1 cells ,and mRNA level was up-regulated by LPS treatment. We also showed expression of resistin mRNA in RAW264.7, a mouse macrophage-like cell line, and its expression was induced by LPS as in human cells. These results suggesting that resistin might be involved in the inflammatory process.
Transfection of resistin-Luciferase reporter gene into RAW264.7 showed that Luciferase activity was significantly up-regulated by LPS treatment. Two DNA fragments, from –1037 to –969 and –968 to –660, were found to be essential for LPS induction of Luciferase activity in 5’ deletion analysis. In this study, we aimed to identify the regulatory elements within the nt –1037 to -969 and nt –968 to –660 of resistin promoter that involved in LPS-induced expression of resistin in RAW264.7 cells.
We constructed various internal deletion (

並列關鍵字

RAW264.7 ； LPS ； octamer ； resistin

參考文獻

姚文雅 (2003) 脂多醣類刺激RAW264.7細胞resistin基因表現增加其分子機制的探討。台灣大學生物化學暨分子生物學研究所碩士論文

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