Myeloid cell leukemia-1 (Mc1-1) is a candidate target gene of hypoxia-inducible factor-1 (HIF-1) in the testis.
Myeloid cell leukemia-1 (Mc1-1) is a candidate target gene of hypoxia-inducible factor-1 (HIF-1) in the testis.
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DOI：
10.1186/1477-7827-10-104
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发表时间：
2012-12-05
期刊：
影响因子：
--
通讯作者：
Karpodinis M
中科院分区：
文献类型：
--
作者：
Palladino MA;Shah A;Tyson R;Horvath J;Dugan C;Karpodinis M
精索扭转可导致睾丸缺血(I)和随后的缺血-再灌注（I/R），引起生殖细胞特异性凋亡。在此之前，我们发现缺氧诱导因子-1 （HIF-1）转录因子在正常和缺血睾丸的间质细胞中含量丰富，是对缺氧生理反应的关键调节因子。我们假设睾丸HIF-1激活抗凋亡靶基因的表达以保护间质细胞免于凋亡。通过对含有缺氧反应元件（HRE, 5′-RCGTG-3′）的睾丸基因的硅分析，发现髓样细胞白血病-1 （Mcl-1）是HIF-1的潜在靶基因。本研究的目的是确定HIF-1在正常缺氧和缺血睾丸中是否显示dna结合活性，以及Mcl-1是否是睾丸HIF-1的靶基因。采用定量酶联免疫吸附试验（ELISA）和电泳迁移转移试验（EMSA）在体外分析睾丸HIF-1 dna结合能力。免疫印迹和免疫组织化学检测MCL-1蛋白表达。通过染色质免疫沉淀（ChIP）分析检测睾丸HIF-1与Mcl-1基因的结合。ELISA和EMSA分析表明，来自常氧和缺血睾丸的HIF-1与DNA的结合同样强烈，这表明HIF-1在常氧睾丸中具有生理作用，而不像大多数组织中HIF-1在常氧条件下被降解，只在缺氧时被激活。MCL-1蛋白在正常缺氧和缺血睾丸中均丰富，并在睾丸间质细胞中表达。与HIF-1表达模式相同，I或I/R和MCL-1与HIF-1α共定位在间质细胞中，MCL-1的稳态水平不受显著影响。使用HIF-1抗体的染色质免疫沉淀（ChIP）分析显示Mcl-1启动子富集的序列。结果表明，与在大多数组织中观察到的不同，HIF-1在正常缺氧和缺血睾丸中都表现出dna结合活性，Mcl-1可能是睾丸HIF-1的关键靶基因，在Leydig细胞的抗凋亡保护中具有潜在的作用。
Spermatic cord torsion can lead to testis ischemia (I) and subsequent ischemia-reperfusion (I/R) causing germ cell-specific apoptosis. Previously, we demonstrated that the hypoxia-inducible factor-1 (HIF-1) transcription factor, a key regulator of physiological responses to hypoxia, is abundant in Leydig cells in normoxic and ischemic testes. We hypothesize that testicular HIF-1 activates the expression of antiapoptotic target genes to protect Leydig cells from apoptosis. In silico analysis of testis genes containing a consensus hypoxia response element (HRE, 5’-RCGTG-3’) identified myeloid cell leukemia-1 (Mcl-1) as a potential HIF-1 target gene. The purpose of this study was to determine whether HIF-1 shows DNA-binding activity in normoxic and ischemic testes and whether Mcl-1 is a target gene of testicular HIF-1. The testicular HIF-1 DNA-binding capacity was analyzed in vitro using a quantitative enzyme-linked immunosorbent assay (ELISA) and electrophoretic mobility shift assays (EMSA). MCL-1 protein expression was evaluated by immunoblot analysis and immunohistochemistry. The binding of testicular HIF-1 to the Mcl-1 gene was examined via chromatin immunoprecipitation (ChIP) analysis. The ELISA and EMSA assays demonstrated that testicular HIF-1 from normoxic and ischemic testes binds DNA equally strongly, suggesting physiological roles for HIF-1 in the normoxic testis, unlike most tissues in which HIF-1 is degraded under normoxic conditions and is only activated by hypoxia. MCL-1 protein was determined to be abundant in both normoxic and ischemic testes and expressed in Leydig cells. In a pattern identical to that of HIF-1 expression, the steady-state levels of MCL-1 were not significantly affected by I or I/R and MCL-1 co-localized with HIF-1α in Leydig cells. Chromatin immunoprecipitation (ChIP) analysis using a HIF-1 antibody revealed sequences enriched for the Mcl-1 promoter. The results demonstrated that, unlike what is observed in most tissues, HIF-1 displays DNA-binding activity in both normoxic and ischemic testes, and Mcl-1 may be a key target gene of testicular HIF-1 with potential roles in the antiapoptotic protection of Leydig cells.
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影响因子：
3.7
作者：
BelAiba, RS;Djordjevic, T;Görlach, A
通讯作者：
Görlach, A
影响因子：
--
作者：
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作者：
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作者：
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通讯作者：
Hermo, L.
影响因子：
64.5
作者：
Semenza GL
通讯作者：
Semenza GL