Microarray analysis of differentiation-specific gene expression during 3T3-L1 adipogenesis
Microarray analysis of differentiation-specific gene expression during 3T3-L1 adipogenesis
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DOI：
10.1016/j.gene.2003.12.012
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发表时间：
2004-03-31
期刊：
影响因子：
3.5
通讯作者：
McGehee, RE
中科院分区：
文献类型：
--
作者：
Burton, GR;Nagarajan, R;McGehee, RE
在细胞分化和发育过程中，人们认识到许多复杂的分子机制以及差异表达基因的精确模式发生在将前体细胞导向给定谱系中。使用基于微阵列的技术，我们研究了3 T3-L1脂肪细胞分化过程中的基因表达。在用标准分化激素诱导剂处理后0、2、8、16、24、48和96 h分离总细胞RNA;胰岛素、地塞米松、异丁基甲基黄嘌呤（IDX）或IDX加阿司他丁A（TsA），一种历史性脱乙酰酶抑制剂和有效的脂肪形成抑制剂。从细胞RNA合成cRNA，并与含有12，488个cDNA/表达序列标签（EST）探针组的高密度Affyphase MG_U74Av2微阵列基因芯片杂交。从仅IDX处理的细胞中，将在整个分化过程中相对于时间0前脂肪细胞未改变或差异表达小于2倍的所有探针组从进一步分析中排除。这种选择导致净1686个转录物，859个在表达中增加，827个在分化中表达降低至少2倍。为了集中于对分化更特异的基因，对IDX加TsA处理的非分化细胞进行相同的分析，并且排除来自仅IDX组的在非分化TsA处理组中表现出相似表达谱的所有探针组，留下仅在分化条件下调节的总共1016个转录物。636这些转录升高至少2倍，380表现出相对于时间0前脂肪细胞的表达减少。这组基因进一步分析，使用分层聚类和自组织地图，并导致许多基因的识别，以前不知道在脂肪细胞分化过程中进行调节。这些基因中的许多可能代表新的脂肪形成介质和脂肪形成的标志物。(C)2004 Elsevier B.V保留所有权利。
During cellular differentiation and development, it is recognized that many complex molecular mechanisms as well as precise patterns of differentially expressed genes occur in directing precursor cells toward a given lineage. Using microarray-based technology, we examined gene expression across the course of 3T3-L1 adipocyte differentiation. Total cellular RNA was isolated at times 0, 2, 8, 16, 24, 48, and 96 h following treatment with either standard hormonal inducers of differentiation; insulin, dexamethasone, isobutylmethylxanthine (IDX), or IDX plus trichostatin A (TsA), a historic deacetylase inhibitor and potent adipogenic inhibitor. cRNA was synthesized from cellular RNA and hybridized to high density Affymetrix MG_U74Av2 microarray gene chips containing 12,488 cDNA/Expressed Sequence Tags (ESTs) probe sets. From the IDX-only treated cells, all probe sets that were either unchanged or differentially expressed less than 2-fold throughout differentiation with respect to time 0 preadipocytes were excluded from further analyses. This selection resulted in a net of 1686 transcripts, 859 were increased in expression, and 827 were decreased in expression at least 2-fold across differentiation. To focus in on genes that were more specific to differentiation, the same analysis was performed on IDX plus TsA-treated non-differentiating cells and all probe sets from the IDX-only group that exhibited similar expression profiles in the non-differentiating TsA-treated group were excluded leaving a total of 1016 transcripts that were regulated only under differentiating conditions. Six hundred and thirty-six of these transcripts were elevated at least 2-fold and 380 exhibited a decrease in expression relative to time 0 preadipocytes. This group of genes was further analyzed using hierarchical clustering and self-organizing maps and resulted in the identification of numerous genes not previously known to be regulated during adipocyte differentiation. Many of these genes may well represent novel adipogenic mediators and markers of adipogenesis. (C) 2004 Elsevier B.V All rights reserved.