Narrowing of the regions of allelic losses of chromosome 1p36 in meningioma tissues by an improved SSCP analysis
Narrowing of the regions of allelic losses of chromosome 1p36 in meningioma tissues by an improved SSCP analysis
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DOI：
10.1002/ijc.23297
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发表时间：
2008-04-15
影响因子：
6.4
通讯作者：
Hayashi, Kenshi
中科院分区：
文献类型：
--
作者：
Guan, Yanlei;Hata, Nobuhiro;Hayashi, Kenshi
在肿瘤组织基因组中定位杂合性缺失(LOH)区域是识别其丢失与肿瘤发生有关的基因的一种实用方法。使用微卫星或单核苷酸多态(SNP)标记的传统杂合性缺失分析需要同时检查肿瘤和匹配的正常DNA。在这里，我们使用单链构象多态(SSCP)分析改进了先前开发的基于SNP的LOH分析，以便即使在没有匹配的正常DNA的情况下，现在也可以准确地估计被正常DNA严重污染的肿瘤样本的LOH。通过与以往的“定量SSCP杂合性丢失检测”(LOQUS-AC)方法的结果比较，我们证明了改进的基于SSCP的杂合性缺失检测方法的可靠性。用LOQUS-AC方法检测LOH与以前的LOQUS方法有很高的一致性(98.1%)。然后，我们应用这种新方法来描述130个脑膜瘤的LOH谱，使用68个SNP(即，平均SNP间间隔441KBP)，这些SNP均匀分布在染色体1p36上。良性脑膜瘤、不典型脑膜瘤和间变性脑膜瘤的LOH频率分别为48.39%、84.62%和100.00%。随后，我们在1p36.11上检测到一个候选的共同杂合性缺失区域，该区域可能含有与脑膜瘤恶性进展相关的肿瘤抑制基因。(C)2007年Wiley-Liss，Inc.
Mapping loss of heterozygosity (LOH) regions in the genomes of tumor tissues is a practical approach for identifying genes whose loss is related to tumorigenesis. Conventional LOH analyses using microsatellite or single nucleotide polymorphism (SNP) markers require the simultaneous examination of tumor- and matched normal-DNA. Here, we improved the previously developed SNP-based LOH assay using single strand conformation polymorphism (SSCP) analysis, so that LOH in tumor samples heavily contaminated with normal DNA can now be precisely estimated, even when matched normal DNA is not available. We demonstrate the reliability of the improved SSCP-based LOH detection method, called the LOH estimation by quantitative SSCP analysis using averaged control (LOQUS-AC), by comparing the results with those of the previous "LOH estimated by quantitative SSCP assay" (LOQUS) method. Using the LOQUS-AC assay, LOH was detected at a high consistency (98.1%) with the previous LOQUS method. We then applied this new method to characterize LOH profiles in 130 meningiomas, using 68 SNPs (i.e., a mean inter-SNP interval of 441 kbp) that are evenly distributed throughout chromosome 1p36. Benign, atypical and anaplastic meningiomas exhibited 1p36 LOH at frequencies of 48.39, 84.62 and 100.00%, respectively, using LOQUS-AC. Subsequently, we detected a candidate common LOH region on 1p36.11 that might harbor tumor suppressor genes related to malignant progression of meningioma. (c) 2007 Wiley-Liss, Inc.