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A hypoxia/radiation dual-sensitive promoter and its transcriptional activity
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 6
- DOI:
- 10.3321/j.issn:1000-2790.2004.06.004
- Key Word:
- 反应元件;生长反应基因-1;低氧;辐射;电离
Abstract： 目的: 构建缺氧/辐射双敏感性启动子,阐明其转录调控特性. 方法: 通过基因重组将缺氧反应元件(HREs)序列插入早期生长反应基因-1(Egr-1)启动子的上游,构成缺氧/辐射双敏感性HRE-Egr启动子及其报告载体pHEL,转染肺癌A549细胞. 在2, 4, 6, 8, 10 Gy照射(γ-射线)后及合并缺氧(氧浓度1, 5, 10, 25, 50 mL/L)时,检测转染细胞内报告基因荧光素酶(luciferase, Luc)表达活性,以及在6 Gy照射合并氧浓度10 mL/L时Luc表达的动态变化. 结果: 成功构建了HRE-Egr启动子及其报告载体,转染A549细胞,发现常氧下HRE-Egr启动子照射后表达活性迅速升高,呈辐射剂量依赖性,与Egr-1启动子相一致. 缺氧时,Egr-1启动子辐射诱导活性明显下降(P<0.05),而HRE-Egr启动子活性则显著升高(P<0.01),同时后者活性持续时间(32 h)明显长于前者(16 h). 结论: HRE-Egr启动子具有辐射/缺氧双重敏感性,在缺氧条件下其辐射诱导活性和持续时间明显增强和延长.
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