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Development of radiation inducible promoter in prostate cancer

Development of radiation inducible promoter in prostate cancer
前列腺癌辐射诱导启动子的开发
批准号:
21791494
负责人:
WATANABE Akihiko
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
项目状态:
已结题
起止时间:
2009 至 2010

项目摘要

项目成果

WATANABE Akihiko的其他基金

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中文摘要
翻译
开发了一个启动子文库,该文库由随机延长转录因子顺式作用元件构建的DNA片段组成,该转录因子可能通过辐射在前列腺癌中激活,并连接到TATA盒序列。在文库的LNCaP细胞中选择了一个对X射线反应最强的启动子,并通过引入随机突变进行了改进。由此产生的启动子被命名为克隆880-8,在X射线照射(X射线照射)下表现出最高的剂量依赖性活性增强。将880-8克隆控制下的重组逆转录病毒感染LNCaP,经10GyX射线照射12h后,荧光素酶活性增加9.12±0.36倍。将感染细胞接种到裸鼠体内,10GyX射线照射12h后,荧光素酶的表达增强了4.27±1.36倍。当另一个携带FCY::Fur基因的重组体在克隆880-8下游感染LNCaP时,经X射线照射后,FCY::Fur的表达增强。与未经X-射线照射的对照相比,5-氟胞嘧啶可提高剂量依赖性的细胞杀伤率。这些结果表明,本研究所采用的方法是构建对刺激反应的启动子的有效方法。这样的启动子可以用于刺激控制的基因治疗。
英文摘要
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.
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会议论文
射線による遺伝子発現制御と前立腺癌の治療応用への検討
放射诱导的基因表达控制及其在前列腺癌治疗中的应用
DOI: --
发表时间: 2010
期刊:
影响因子: --
作者: [森井章裕, 渡部明彦, 他]
通讯作者: 他
前立腺癌細胞株における放射線遺伝子治療用人工プロモーターの開発
前列腺癌细胞系放射基因治疗人工启动子的开发
DOI: --
发表时间: 2011
期刊:
影响因子: --
作者: [森井章裕, 渡部明彦, 他]
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放射線による遺伝子発現制御と前立腺癌の治療応用への検討
放射控制基因表达的思考及其在前列腺癌治疗中的应用
DOI: --
发表时间: 2010
期刊:
影响因子: --
作者: [森井章裕、渡部明彦, 他]
通讯作者: 他
前立腺癌細胞株において放射線に応答する人工プロモーターの改良
前列腺癌细胞系中响应辐射的人工启动子的改进
DOI: --
发表时间: 2009
期刊:
影响因子: --
作者: [森井章裕, 渡部明彦, 他]
通讯作者: 他
共 6 条
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