Mutations in the Circadian Gene CLOCK in Colorectal Cancer
Mutations in the Circadian Gene CLOCK in Colorectal Cancer
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DOI：
10.1158/1541-7786.mcr-10-0086
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发表时间：
2010-07-01
影响因子：
5.2
通讯作者：
Aaltonen, Lauri A.
中科院分区：
文献类型：
--
作者：
Alhopuro, Pia;Bjorklund, Mikael;Aaltonen, Lauri A.
生物钟调节生理过程的每日变化。CLOCK在昼夜节律装置中起调节器的作用，控制其他时钟基因（包括PER 1）的表达。时钟基因与癌症相关的功能有关;在这项工作中，我们研究了CLOCK作为微卫星不稳定结直肠癌体细胞突变的可能靶点。结合微阵列基因表达数据和公开的基因序列信息，我们确定CLOCK为790个推定的新型微卫星不稳定性（MSI）靶基因之一。对101例结直肠癌（CRC）的CLOCK基因编码微卫星进行了测序。通过稳定表达GST-CLOCK或谷胱甘肽S-转移酶空载体并通过使用流式细胞术的DNA含量分析测试UV诱导的细胞凋亡和辐射的影响，在缺乏野生型CLOCK的LS 180细胞中研究恢复CLOCK表达的效果。通过使用ChIP-seq搜索推定的新型CLOCK靶基因。CLOCK突变发生在53%的MSI CRC中。在双等位基因CLOCK失活的细胞中恢复CLOCK表达导致对UV诱导的细胞凋亡的保护和响应于电离辐射的G2-M期阻滞的减少。使用ChIP-Seq，在DNA损伤基因p21、NBR 1、BRCA 1和RAD 50附近鉴定了新的CLOCK结合元件。CLOCK在癌症中被证明是突变的，对DNA损伤的反应改变提供了一种可能的肿瘤发生机制。Mol Cancer Res; 8（7）; 952-60. (C)2010年AACR。
The circadian clock regulates daily variations in physiologic processes. CLOCK acts as a regulator in the circadian apparatus controlling the expression of other clock genes, including PER1. Clock genes have been implicated in cancer-related functions; in this work, we investigated CLOCK as a possible target of somatic mutations in microsatellite unstable colorectal cancers. Combining microarray gene expression data and public gene sequence information, we identified CLOCK as 1 of 790 putative novel microsatellite instability (MSI) target genes. A total of 101 MSI colorectal carcinomas (CRC) were sequenced for a coding microsatellite in CLOCK. The effect of restoring CLOCK expression was studied in LS180 cells lacking wild-type CLOCK by stably expressing GST-CLOCK or glutathione S-transferase empty vector and testing the effects of UV-induced apoptosis and radiation by DNA content analysis using flow cytometry. Putative novel CLOCK target genes were searched by using ChIP-seq. CLOCK mutations occurred in 53% of MSI CRCs. Restoring CLOCK expression in cells with biallelic CLOCK inactivation resulted in protection against UV-induced apoptosis and decreased G2-M arrest in response to ionizing radiation. Using ChIP-Seq, novel CLOCK-binding elements were identified near DNA damage genes p21, NBR1, BRCA1, and RAD50. CLOCK is shown to be mutated in cancer, and altered response to DNA damage provides one plausible mechanism of tumorigenesis. Mol Cancer Res; 8(7); 952-60. (C) 2010 AACR.