Harmonized microarray/mutation scanning analysis of TP53 mutations in undissected colorectal tumors
Harmonized microarray/mutation scanning analysis of TP53 mutations in undissected colorectal tumors
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DOI：
10.1002/humu.20069
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发表时间：
2004-01-01
期刊：
影响因子：
3.9
通讯作者：
Barany, F
中科院分区：
文献类型：
--
作者：
Favis, R;Huang, JM;Barany, F
TP 53（p53）的突变状态和特异性突变已被证明会影响肿瘤预后和对治疗的反应。由于正常DNA可干扰突变序列的检测，因此实体瘤的分子谱分析受到浸润性野生型细胞的混淆。我们的目的是在138例I-IV期结直肠腺癌和肝转移瘤中鉴定TP 53突变，而不首先通过显微切片富集肿瘤细胞。为了实现这一点，我们开发了一个协调的协议，涉及多重聚合酶链反应/连接酶检测反应（PCR/LDR）与通用DNA微阵列分析和核酸内切酶V/连接酶突变扫描。使用双脱氧测序验证序列。协调方案检测到所有66种突变。双脱氧测序使用自动阅读检测到66个突变中的41个（62%），使用手动阅读检测到66个突变中的59个（89%）。将TP 53数据库（http：//www.example.com）中的结肠癌条目与本研究中报道的结果进行比较的数据分析显示，突变的分布和突变事件是相当的。p53.curie.fr (C)2004威利-利斯公司
Both the mutational status and the specific mutation of TP53 (p53) have been shown to impact both tumor prognosis and response to therapies. Molecular profiling of solid tumors is confounded by infiltrating wild-type cells, since normal DNA can interfere with detection of mutant sequences. Our objective was to identify TP53, mutations in 138 stage I-IV colorectal adenocarcinomas and liver metastases without first enriching for tumor cells by microdis section. To achieve this, we developed a harmonized protocol involving multiplex polymerase chain reaction/ligase detection reaction (PCR/LDR) with Universal DNA microarray analysis and endonuclease V/Ligase mutation scanning. Sequences were verified using dideoxy sequencing. The harmonized protocol detected all 66 mutations. Dideoxy sequencing detected 41 out of 66 mutations (62%) using automated reading, and 59 out of 66 mutations (89%) with manual reading. Data analysis comparing colon cancer entries in the TP53 database (http://p53.curie.fr) with the results reported in this study showed that distribution of mutations and the mutational events were comparable. (C) 2004 Wiley-Liss, Inc.