Sucrose-induced vacuolation results in increased expression of cholesterol biosynthesis and lysosomal genes
Sucrose-induced vacuolation results in increased expression of cholesterol biosynthesis and lysosomal genes
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DOI：
10.1016/j.yexcr.2003.09.003
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发表时间：
2004-01-01
影响因子：
3.7
通讯作者：
Thoene, JG
中科院分区：
文献类型：
--
作者：
Helip-Wooley, A;Thoene, JG
在高浓度蔗糖存在下培养的哺乳动物细胞表现出大的、相透明的、肿胀的空泡。三个正常的人成纤维细胞系暴露于100 mM的蔗糖24小时表现出增加表达的溶酶体，胞内囊泡运输，胆固醇生物合成，和脂肪酸代谢基因。胆固醇生物合成途径的大多数步骤被上调，包括HMG CoA还原酶，其催化胆固醇生物合成的限速步骤。溶酶体基因神经氨酸酶、CLN 3和CLCN 5以及小GTP结合蛋白Rab 7 L1和Ar 17也增加。Rab 7 L1-GFP融合蛋白在人成纤维细胞中过表达，并且被证明主要定位于高尔基体，并且在一些细胞中定位于核周区域中的膜边界囊泡。增加水平的转录因子C/EBP被发现在细胞核提取物中暴露于蔗糖12小时，相对于匹配的控制表明调节基因表达后蔗糖诱导的空泡化可能是协调的，至少部分地，由转录因子C/EBP。蔗糖诱导的空泡化是研究溶酶体基因表达调控和生物发生的一个有用模型。(C)2003年爱思唯尔公司All rights reserved.
Mammalian cells cultured in the presence of high concentrations of sucrose demonstrate large, phase-lucent, osmotically swollen vacuoles. Three normal human fibroblast cell lines exposed to 100 mM of sucrose for 24 h demonstrated increased expression of lysosomal, intracellular vesicle trafficking, cholesterol biosynthesis, and fatty acid metabolism genes. Most steps of the cholesterol biosynthesis pathway were upregulated including HMG CoA reductase, which catalyzes the rate-limiting step of cholesterol biosynthesis. The lysosomal genes neuraminidase, CLN3, and CLCN5 and the small GTP-binding proteins Rab7L1 and Ar17 were also increased. A Rab7L1-GFP fusion protein was overexpressed in human fibroblasts and was demonstrated to localize primarily to the Golgi apparatus, and in some cells to the membranes bounding vesicles in the perinuclear region. Increased levels of the transcription factor C/EBP were found in nuclear extracts from cells exposed to sucrose for 12 h, relative to matched controls suggesting regulation of gene expression following sucrose-induced vacuolation may be coordinated, at least in part, by the transcription factor C/EBP. Sucrose-induced vacuolation is a useful model in which to study the regulation of lysosomal gene expression and biogenesis. (C) 2003 Elsevier Inc. All rights reserved.