Development of radiation inducible promoter in prostate cancer
前列腺癌辐射诱导启动子的开发
基本信息
- 批准号:21791494
- 负责人:
- 金额:$ 2.5万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Young Scientists (B)
- 财政年份:2009
- 资助国家:日本
- 起止时间:2009 至 2010
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
A promoter library was developed that is composed of DNA fragments constructed by randomly elongating cis-acting elements of transcription factors presumably activated in prostate cancer by radiation, and linking to the TATA box sequence. One promoter with the strongest reactivity to X-ray in LNCap cells of the library was chosen and improved by the introduction of random mutations. A resultant promoter was designated clone 880-8 showing the highest dose-dependent activity enhancement with X-ray irradiation (X-irradiation). A recombinant retrovirus expressing the luciferase gene under the control of clone 880-8 was infected to LNCap that showed 9.12 ± 0.36-fold enhancement of luciferase activity 12 h after X-irradiation at 10 Gy. When the infected cells were inoculated onto nude mice, enhancement of luciferase expression was 4.27 ± 1.36-fold 12 h after X-irradiation at 10 Gy. When LNCap was infected with another recombinant carrying the fcy::fur gene downstream to clone 880-8, fcy::fur expression was enhanced by X-irradiation. It was also shown to increase dose-dependent cell killing ratio with 5-fluorocytosine compared to a counterpart without X-irradiation. These results suggest that the method employed in this study is effective to construct a promoter responsive to stimulation. Such promoters can be utilized for stimulation controlled-gene therapies.
开发了一个启动子库,该库由通过辐射通过辐射中前列腺癌中的转录因子的顺式作用元件的随机拉长构建的DNA片段组成,并链接到TATA盒序列。选择了一个对库中LNCAP细胞中X射线反应性最强的启动子,并通过引入随机突变进行了改进。将最终的启动子指定为克隆880-8,显示出最高剂量依赖性活性增强,并通过X射线照射(X-Irradiation)。将克隆880-8控制下表达荧光素酶基因的重组逆转录病毒感染到LNCAP,在10 Gy X射线后12小时,荧光素酶活性增强了9.12±0.36倍。当感染细胞被接种到裸鼠上时,在10 Gy时X-辐照后,荧光素酶表达的增强为4.27±1.36倍。当LNCAP被另一种重组携带FCY的重组感染时,将FCY :: Fur Gene下游到克隆880-8时,FCY :: fcy :: fur ::通过X-irraradiatiation增强了毛皮表达。与没有X辐射的对应物相比,它还显示出5-氟环胞嘧啶的剂量依赖性细胞杀伤比。这些结果表明,本研究中采用的方法有效地构建了响应刺激的启动子。此类启动子可以用于刺激控制的基因疗法。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Construction of promoter induced by radiation for human prostate cancer cells.
人前列腺癌细胞辐射诱导启动子的构建。
- DOI:
- 发表时间:2009
- 期刊:
- 影响因子:0
- 作者:Akihiro Morii;Akihiko Watanabe;et al.
- 通讯作者:et al.
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WATANABE Akihiko其他文献
WATANABE Akihiko的其他文献
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