AHR, a novel acute hypoxia-response sequence, drives reporter gene expression under hypoxia in vitro and in vivo
AHR, a novel acute hypoxia-response sequence, drives reporter gene expression under hypoxia in vitro and in vivo
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DOI：
10.1042/cbi20100290
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发表时间：
2011-01-01
影响因子：
3.9
通讯作者：
Ninomiya, Yoshifumi
中科院分区：
文献类型：
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作者：
Cilek, Mehmet Zeynel;Hirohata, Satoshi;Ninomiya, Yoshifumi
ADAMTS1（一种具有血小板反应蛋白基序 1 的解整合素和金属蛋白酶）是一种早期直接基因。我们之前报道过 ADAMTS1 是由缺氧强烈诱导的。在本研究中，我们研究了 ADAMTS1 启动子驱动的报告信号是否可以通过急性缺氧检测到。我们构建了ADAMTS1启动子控制下的GFP（绿色荧光蛋白）表达载体[AHR（急性缺氧反应序列）-GFP]，并将其与CMV（巨细胞病毒启动子-OFF）控制下的组成型GFP表达载体进行了比较。我们转导AHR-GFP并检查在急性缺氧下是否可以检测到OFF信号。常氧条件下转导人脐静脉[HUVEC（人脐静脉内皮细胞）]时，GFP信号很少，而CMV-GFP则显示出相当多的GFP信号。当HUVEC受到缺氧刺激时，AHR-GFP基因的OFF信号在缺氧条件下被诱导。值得注意的是，OFF 信号在缺氧条件下 3 小时达到峰值。在缺血后肢模型中，转导的 AHR-GFP 显示 GFP 信号的缺氧诱导。总之，我们证明AHR系统通过急性缺氧诱导报告基因表达，并且其诱导是短暂的。这是第一份展示独特的急性缺氧激活基因表达系统的报告。
ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1) is an early immediate gene. We have previously reported that ADAMTS1 was strongly induced by hypoxia. In this study, we investigated whether ADAMTS1 promoter-driven reporter signal is detectable by acute hypoxia. We constructed the GFP (green fluorescent protein) expression vector [AHR (acute hypoxia-response sequence)-GFP] under the control of ADAMTS1 promoter and compared it with the constitutive GFP-expressing vector under the control of CMV (cytomegalovirus promoter-OFF). We transduced AHR-GFP and examined whether OFF signals can be detected under the acute hypoxia. When the human umbilical vein [HUVEC (human umbilical vein endothelial cells)] was transduced under normoxia, there were few GFP signals, while CMV-GFP showed considerable GFP signals. When HUVEC was stimulated with hypoxia, OFF signals from AHR-GFP gene were induced under hypoxic conditions. Notably, the OFF signals peaked at 3 h under hypoxia. In ischaemic hind limb model, transduced AHR-GFP showed hypoxic induction of GFP signals. In summary, we have demonstrated that the AHR system induced the reporter gene expression by acute hypoxia, and its induction is transient. This is the first report showing the unique acute hypoxia-activated gene expression system.