Afr2 regulation occurs cell-autonomously in vitro but is not conferred on episomal DNA in transient assays.
Afr2 regulation occurs cell-autonomously in vitro but is not conferred on episomal DNA in transient assays.
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Afr2 调节在体外细胞自主发生，但在瞬时测定中并未赋予附加型 DNA。
DOI：
10.1089/dna.2005.24.189
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Feuerman,MiriamH
中科院分区：
文献类型：
--
作者：
Park,JamesK;Feuerman,MiriamH
癌胚抗原如甲胎蛋白（AFP）在再生肝脏中表达。肝再生过程中AFP基因的表达水平受非连锁的常染色体基因--甲胎蛋白调节因子2（Afr 2）的调节。C3 H/HeJ（Afr 2 A/A）小鼠表达的AFP水平比C57 BL/6 J（Afr 2 B/B）小鼠高10倍。在这里，我们表明，从C3 H/HeJ和C57 BL/6 J小鼠分离的原代肝细胞表现出内源性AFP基因的差异表达，这是由于Afr 2基因位点和指示的cellaureus机制。我们发现，theAfr 2-ResponseElement（ARE），AFP转录起始位点上游1010和838碱基对之间，没有调节报告基因的表达在Hep G2，Hep 3B，Hepa 1.6和HeLa细胞系的转染试验。瞬时转染的原代肝细胞中的报告基因表达也是ARE-独立的。最后，CCl 4诱导的肝再生后，通过基于流体动力学的转染向C3 H/HeJ和C57 BL/6 J小鼠的肝脏递送的报告构建体的基因表达是ARE独立的。总之，ARE依赖性转录没有发现在三个不同的系统，其中两个保留内源性AFP基因的调节，表明ARE可能不作为一个简单的转录因子识别位点的瞬时测定。
Oncofetal antigens such as alpha-fetoprotein (AFP) are expressed in regenerating liver. The level of AFP gene expression during liver regeneration is regulated by the unlinked, autosomal gene,Alpha-fetoprotein regulator 2 (Afr2). C3H/HeJ (Afr2A/A) mice express 10-fold higher levels of AFP than C57BL/6J (Afr2B/B) mice. Here we show that primary hepatocytes isolated from C3H/HeJ and C57BL/6J mice exhibit differential expression of the endogenous AFP gene, which was attributed to the Afr2 gene locus and indicative of a cellautonomous mechanism. We show that theAfr2-ResponseElement (ARE), between 1010 and 838 base pairs upstream of the AFP transcriptional start site, did not modulate reporter gene expression in transfection assays of Hep G2, Hep 3B, Hepa 1.6, and HeLa cell lines. Reporter gene expression in transiently transfected primary hepatocytes was also ARE-independent. Finally, gene expression from reporter constructs delivered by hydrodynamics-based transfection to the livers of C3H/HeJ and C57BL/6J mice after CCl4-induced liver regeneration was ARE-independent. In conclusion, ARE-dependent transcription was not found in transient assays performed in three different systems, two of which retained regulation of the endogenous AFP gene, suggesting that the ARE may not function as a simple transcription factor recognition site.