Identification and comparative analysis of human colonocyte short-chain fatty acid response genes
Identification and comparative analysis of human colonocyte short-chain fatty acid response genes
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DOI：
10.1016/s1091-255x(00)80093-1
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发表时间：
2000-09-01
影响因子：
3.2
通讯作者：
Rothberg, BEG
中科院分区：
文献类型：
--
作者：
Basson, MD;Liu, YW;Rothberg, BEG
纤维发酵过程中产生的短链脂肪酸（SCFA）丁酸酯、丙酸酯和乙酸酯促进结肠分化，并可逆转或抑制肿瘤进展。我们试图确定负责结肠细胞上SCFA活性的候选基因，并比较独立SCFA的相对活性。从对照Caco-2细胞和用等摩尔丁酸盐、丙酸盐和乙酸盐处理的细胞中分离的polyA（+）mRNA产生cDNA。应用了GeneCalling，一种与DNA序列数据库查找相关的基于限制的差异mRNA表达平台。分析了总共30，000个个体基因序列在三种SCFA中的差异表达。将对应于癌症相关基因的差异表达峰分离、测序并交叉引用到GenBank人类数据库。通过寡核苷酸中毒独立确认基因身份。超过1000个基因片段被鉴定为丁酸盐在表达方面受到实质性调节。丁酸盐的效果最明显，乙酸盐的效果最差。选择用于进一步研究的五个片段进行了完全测序，并证明与人类簇蛋白，淀粉样蛋白受体样蛋白2和尾部同源框2蛋白序列100%同源，以前不知道由SCFAs调制。在每种情况下，观察到研究的三种SCFA的效力顺序相似。常见的SCFA似乎发挥不同的作用。这项研究表明SCFA反应在分子水平上的多样性，并有助于确定膳食纤维生物活性中重要的基因。
Short-chain fatty acids (SCFAs) butyrate, propionate, and acetate produced during fiber fermentation promote colonic differentiation and can reverse or suppress neoplastic progression. We sought to identify candidate genes responsible for SCFA activity on colonocytes and to compare the relative activities of independent SCFAs. cDNA was generated from polyA(+) mRNA isolated from control Caco-2 cells and cells treated with equimolar butyrate, propionate, and acetate. GeneCalling, a restriction-based differential mRNA expression platform linked to a DNA sequence database lookup, was applied. A total of 30,000 individual genetic sequences were analzyed for differential expression among the three SCFAs. Differentially expressed peaks corresponding to cancer-related genes were isolated, sequenced, and cross-referenced to the GenBank human database. Gene identities were independently confirmed by oligonucleotide poisoning. More than 1000 gene fragments were identified as being substantially modulated in expression by butyrate. Butyrate tended to have the most pronounced effects and acetate the least. Five fragments selected for further study were fully sequenced and proved 100% homologous with human sequences for clusterin, amyloid precursor-like protein 2, and caudal homeobox 2 protein, not previously known to be modulated by SCFAs. In each case, a similar order of potency for the three SCFAs studied was observed. The common SCFAs appear to exert different effects. This study suggests the diversity of the SCFA response at the molecular level and facilitates identifying genes important in the biologic activity of dietary fiber.