四氢生物蝶呤对小鼠成纤维细胞糖代谢的作用机制

发布时间：2018-01-15 11:05

本文关键词：四氢生物蝶呤对小鼠成纤维细胞糖代谢的作用机制　出处：《华侨大学》2014年硕士论文　论文类型：学位论文

【摘要】：与正常分化的细胞不同，多数增殖细胞和肿瘤细胞通过有氧糖酵解的方式为细胞生理过程提供能量，即“Warburg效应”。这种快速有效的代谢重编程满足了增殖细胞对生物大分子合成和能量需求。此外，乳酸的大量排出改变了细胞外的微环境，为肿瘤细胞的存活、生长和侵袭创造了有利条件。四氢生物蝶呤（BH4）是芳香族氨基酸羟化酶和一氧化氮合酶的辅因子。BH4有两种合成途径：从头合成和补救合成。其中GTPCH是从头合成中的限速酶，墨蝶呤是补救合成中的底物。基因芯片结果显示，，GCH基因在结肠癌肝脏转移的基质成纤维细胞中有显著性的高表达。研究表明四氢生物蝶呤在促进血管新生和肿瘤细胞增殖中有重要作用。本文利用已经构建的两种小鼠成纤维细胞为模型，GTPCH表达组细胞命名为“GCH-tet-off”，空载体转染组命名为“Tet-off-EV”。通过外源性添加墨蝶呤和调控GTPCH表达探究BH4的合成对这两种细胞糖代谢以及它们成瘤性的作用。结果显示BH4能够增加葡萄糖利用率和乳酸产量，上调LDHA、PDK1、MCT4和CAⅨ表达。BH4的合成能激活PI3K/Akt/GSK3β信号通路。此外，BH4能诱导成纤维细胞发生致瘤性的转变。这些结果表明BH4促进小鼠成纤维细胞有氧糖酵解代谢表型的改变。
[Abstract]:Unlike normal differentiated cells , most of the proliferating cells and tumor cells provide energy , i.e . , the " Warburg effect " , through an aerobic glycolytic process that satisfies the synthesis and energy requirements of proliferating cells for the biosynthesis and energy of biological macromolecules . In addition , a large amount of lactic acid efflux changes the microenvironment outside the cells , creating favorable conditions for survival , growth and invasion of tumor cells . Tetrahydrobiopterin ( BH 4 ) is a cofactor of aromatic amino acid hydroxylase and nitric oxide synthase . There are two synthetic pathways : de novo synthesis and salvage synthesis . The GTPCH is a restriction enzyme in de novo synthesis . The gene chip shows significant high expression of GCH gene in stromal fibroblasts transferred from colon cancer liver . Studies have shown that tetrahydrobiopterin plays an important role in promoting angiogenesis and tumor cell proliferation . In this paper , two kinds of mouse fibroblast cells have been constructed , GTPCH expression group cells were named " GCH - tet - off " , and the empty vector transfection group was named " GCH - off - EV " . The results showed that BH could increase the glucose utilization rate and lactate production , up - regulate the expression of LDHA , PDK1 , MCT4 and CA IX . The synthesis of BH could activate the pathway of PI3尾 signaling .

【学位授予单位】：华侨大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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