Department of Toxicology and Cancer Biology, University of Kentucky, Lexington, KY, USA.
Methods Mol Biol. 2023;2701:231-242. doi: 10.1007/978-1-0716-3373-1_15.
Cells experience increased genome instability through the course of disease development including cancer initiation and progression. Point mutations, insertion/deletions, translocations, and amplifications of both coding and noncoding regions all contribute to cancer phenotypes. Copy number variation (CNV), i.e., changes of the number of copies of nuclear DNA, occurs in the genome of even normal somatic cells. Studies to understand the effects of CNV on tumor development, especially aspects concerning tumor aggressiveness and the influence on outcomes of therapeutic modalities, have been reignited by the breakthrough technologies of the molecular genomics. This section discusses the significance of analyzing CNVs that cause simultaneous increase/decrease of clusters of genes, using the expression profile of XRCC1 with its neighbor genes LIG1, PNKP, and POLD1 as an example. Methods for CNV assay at the individual gene level on formalin-fixed, paraffin-embedded (FFPE) tissues using the NanoString nCounter technology will then be described.
在疾病发展过程中，包括癌症的起始和进展，细胞会经历基因组不稳定性的增加。点突变、插入/缺失、易位和编码及非编码区域的扩增都会导致癌症表型。拷贝数变异（CNV），即核 DNA 拷贝数的变化，甚至在正常体细胞的基因组中也会发生。通过分子基因组学的突破性技术，对 CNV 对肿瘤发展的影响，特别是对肿瘤侵袭性和对治疗方式结果的影响的研究，重新引起了关注。本节将以 XRCC1 及其相邻基因 LIG1、PNKP 和 POLD1 的表达谱为例，讨论分析同时导致基因簇增加/减少的 CNVs 的意义。然后将描述使用 NanoString nCounter 技术在福尔马林固定、石蜡包埋（FFPE）组织上进行单个基因水平的 CNV 检测的方法。
