RETINOIC ACID PREVENTS DOWN-REGULATION OF RAS RECISION GENE LYSYL OXIDASE EARLY IN ADIPOCYTE DIFFERENTIATION
RETINOIC ACID PREVENTS DOWN-REGULATION OF RAS RECISION GENE LYSYL OXIDASE EARLY IN ADIPOCYTE DIFFERENTIATION
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DOI：
10.1046/j.1432-0436.1994.5810047.x
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发表时间：
1994-11-01
期刊：
影响因子：
2.9
通讯作者：
LAZAR, MA
中科院分区：
文献类型：
--
作者：
DIMACULANGAN, DD;CHAWLA, A;LAZAR, MA
3 T3-L1细胞向脂肪细胞分化是一个复杂的过程，维甲酸（RA）可抑制3 T3-L1细胞向脂肪细胞分化。由于RA的核受体直接调节基因表达的行为，我们假设，在这个系统中的RA行动的主要目标是在脂肪转化的早期调控的基因。在这项研究中，我们展示了使用差异显示技术来寻找早期事件的脂肪承诺是敏感的RA。一个mRNA的基础上确定其RA依赖的基因表达后24小时开始的标准分化方案。分子克隆的cDNA显示它是相同的ras recision基因（rrg），赖氨酰氧化酶。事实上，两种mRNA相同的赖氨酰氧化酶探针识别的前脂肪细胞中表达和tandemly抑制与24小时的暴露分化条件。赖氨酰氧化酶活性在分化细胞的培养基中同样降低。RA完全阻断了分化相关的rrg/赖氨酰氧化酶基因表达的减少，尽管RA对未暴露于分化条件的细胞中的rrg/赖氨酰氧化酶表达没有独立的刺激作用。因此，差异显示已被成功地用于识别rrg/赖氨酰氧化酶作为早期标志物的脂肪转化，是响应于RA。
Adipocyte differentiation of 3T3-L1 cells is a complex process which is inhibited by retinoic acid (RA). Since RA acts by nuclear receptors which directly regulate gene expression, we postulate that the primary targets of RA action in this system are genes which are regulated early in adipose conversion. In this study, we demonstrate the use of the differential display technique to search for early events in adipose commitment which are sensitive to RA. A mRNA was identified on the basis of its RA-dependent gene expression 24 h after initiation of a standard differentiation protocol. Molecular cloning of the cDNA revealed it to be identical to the ras recision gene (rrg), for lysyl oxidase. Indeed, two mRNAs identical to those recognized by lysyl oxidase probes were expressed in preadipocytes and tandemly repressed with 24 h of exposure to differentiation conditions. Lysyl oxidase activity was similarly reduced in the media of differentiated cells. RA completely blocked the differ-entiation-related reduction in rrg/lysyl oxidase gene expression, although RA had no independent stimulatory effect on rrg/lysyl oxidase expression in cells not exposed to differentiating conditions. Thus, differential display has been successfully used to identify rrg/lysyl oxidase as an early marker for adipose conversion that is responsive to RA.