A fast, simple method for screening radiation susceptibility genes by RNA interference
A fast, simple method for screening radiation susceptibility genes by RNA interference
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DOI：
10.1016/j.bbrc.2005.06.047
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发表时间：
2005-08-12
影响因子：
3.1
通讯作者：
Harada, Y
中科院分区：
文献类型：
--
作者：
Tsuji, AB;Sudo, H;Harada, Y
放射治疗可能对某些癌症患者的正常组织造成不可接受的损伤。为了了解辐射诱导的生理反应的分子机制，并能够预测个体患者正常组织的辐射敏感性，重要的是要确定一套全面的基因负责辐射敏感性。我们已经开发了一个简单而快速的96孔筛选协议，使用细胞增殖试验和RNA干扰，以确定与辐射敏感性相关的基因。我们使用96孔板评估了基于alamarBlue、BrdU和磺酰罗丹明B的细胞增殖试验的性能。每个增殖试验检测已知的辐射敏感性基因，PRKDC。在使用28个shRNA载体的试验筛选中，鉴定了另一个已知基因CDKN 1A和一个新的辐射敏感基因ATP 5G3。我们的研究结果表明，这种方法可能是有用的大规模筛选设计，以确定新的辐射易感基因。(c)2005年爱思唯尔公司All rights reserved.
Radiotherapy can cause unacceptable levels of damage to normal tissues in some cancer patients. To understand the molecular mechanisms underlying radiation-induced physiological responses, and to be able to predict the radiation susceptibility of normal tissues in individual patients, it is important to identify a comprehensive set of genes responsible for radiation susceptibility. We have developed a simple and rapid 96-well screening protocol using cell proliferation assays and RNA interference to identify genes associated with radiation susceptibility. We evaluated the performance of alamarBlue-, BrdU-, and sulforhodamine B-based cell proliferation assays using the 96-well format. Each proliferation assay detected the known radiation susceptibility gene, PRKDC. In a trial screen using 28 shRNA vectors, another known gene, CDKN1A, and one new radiation susceptibility gene, ATP5G3, were identified. Our results indicate that this method may be useful for large-scale screens designed to identify novel radiation susceptibility genes. (c) 2005 Elsevier Inc. All rights reserved.