Elahi Eiram, Suraweera Nirosha, Volikos Emmanouil, Haines Jackie, Brown Natalie, Davidson Gerovie, Churchman Mike, Ilyas Mohammed, Tomlinson Ian, Silver Andrew
Colorectal Cancer Genetics, Institute for Cell and Molecular Sciences, Barts and The London, Queen Mary's School of Medicine and Dentistry, London, United Kingdom.
PLoS One. 2009;4(2):e4388. doi: 10.1371/journal.pone.0004388. Epub 2009 Feb 5.
Ionising radiation is a carcinogen capable of inducing tumours, including colorectal cancer, in both humans and animals. By backcrossing a recombinant line of Apc(Min/+) mice to the inbred BALB/c mouse strain, which is unusually sensitive to radiation-induced tumour development, we obtained panels of 2Gy-irradiated and sham-irradiated N2 Apc(Min/+) mice for genotyping with a genome-wide panel of microsatellites at approximately 15 cM density and phenotyping by counting adenomas in the small intestine. Interval and composite interval mapping along with permutation testing identified five significant susceptibility quantitative trait loci (QTLs) responsible for radiation induced tumour multiplicity in the small intestine. These were defined as Mom (Modifier of Min) radiation-induced polyposis (Mrip1-5) on chromosome 2 (log of odds, LOD 2.8, p = 0.0003), two regions within chromosome 5 (LOD 5.2, p<0.00001, 6.2, p<0.00001) and two regions within chromosome 16 respectively (LOD 4.1, p = 4x10(-5), 4.8, p<0.00001). Suggestive QTLs were found for sham-irradiated mice on chromosomes 3, 6 and 13 (LOD 1.7, 1.5 and 2.0 respectively; p<0.005). Genes containing BALB/c specific non-synonymous polymorphisms were identified within Mrip regions and prediction programming used to locate potentially functional polymorphisms. Our study locates the QTL regions responsible for increased radiation-induced intestinal tumorigenesis in Apc(Min/+) mice and identifies candidate genes with predicted functional polymorphisms that are involved in spindle checkpoint and chromosomal stability (Bub1b, Casc5, and Bub1), DNA repair (Recc1 and Prkdc) or inflammation (Duox2, Itgb2l and Cxcl5). Our study demonstrates use of in silico analysis in candidate gene identification as a way of reducing large-scale backcross breeding programmes.
电离辐射是一种致癌物,能够在人类和动物中诱发肿瘤,包括结直肠癌。通过将Apc(Min/+)小鼠的重组品系与对辐射诱导的肿瘤发展异常敏感的近交BALB/c小鼠品系进行回交,我们获得了经2Gy照射和假照射的N2 Apc(Min/+)小鼠组,用于使用密度约为15 cM的全基因组微卫星面板进行基因分型,并通过计数小肠中的腺瘤进行表型分析。区间和复合区间作图以及置换检验确定了五个显著的易感性数量性状基因座(QTL),它们负责小肠中辐射诱导的肿瘤多样性。这些被定义为2号染色体上的Mom(Min修饰因子)辐射诱导的息肉病(Mrip1-5)(优势对数,LOD 2.8,p = 0.0003),5号染色体上的两个区域(LOD 5.2,p<0.00001,6.2,p<0.00001)以及16号染色体上的两个区域(LOD 4.1,p = 4x10(-5),4.8,p<0.00001)。在3号、6号和13号染色体上发现了假照射小鼠的提示性QTL(分别为LOD 1.7、1.5和2.0;p<0.005)。在Mrip区域内鉴定出含有BALB/c特异性非同义多态性的基因,并使用预测程序来定位潜在的功能性多态性。我们的研究定位了导致Apc(Min/+)小鼠辐射诱导的肠道肿瘤发生增加的QTL区域,并鉴定了具有预测功能多态性的候选基因,这些基因参与纺锤体检查点和染色体稳定性(Bub1b、Casc5和Bub1)、DNA修复(Recc1和Prkdc)或炎症(Duox2、Itgb2l和Cxcl5)。我们的研究证明了在候选基因鉴定中使用计算机分析作为减少大规模回交育种计划的一种方法。