IR@PKUHSC  > 北京大学临床肿瘤学院
学科主题临床医学
High throughput detection of microsatellite instability by denaturing high-performance liquid chromatography
Pan, K; Liu, W; Lu, YY; Zhang, L; Li, Z; Lu, WL; Thibodeau, SN; You, WC
关键词microsatellite instability BAT25 BAT26 DHPLC gastric cancer MLH1 MLH2 cancer
刊名HUMAN MUTATION
2003-11-01
DOI10.1002/humu.10271
22期:5页:388-394
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Genetics & Heredity
研究领域[WOS]Genetics & Heredity
关键词[WOS]NONPOLYPOSIS COLORECTAL-CANCER ; SINGLE NUCLEOTIDE POLYMORPHISMS ; REPLICATION ERROR PHENOTYPE ; SOMATIC MUTATIONS ; MISMATCH REPAIR ; TUMORS ; CARCINOMAS ; GERMLINE ; BAT-26 ; DHPLC
英文摘要

Microsatellite instability (MSI) is a hallmark of the DNA replication error phenotype, due to the inactivation of mismatch repair genes. MSI has been implicated in colon and many other gastrointestinal cancers. MSI usually can be analyzed by PCR amplification of microsatellite markers followed by electrophoresis and detected using autoradiography or fluorescence techniques. We report here a novel method for high throughput detection of MSI using denaturing high performance liquid chromatography (DHPLC). Amplification of two mononucleotide markers (BAT25 and BAT26) by polymerase chain reaction (PCR) is followed by DHPLC analysis to display alteration in the length of repetitive sequences. These two markers were tested in 84 colorectal cancer samples confirmed to be 44 MSI-H and 40 MSI-L or MSS, previously defined by multiple microsatellite markers and/or by immunohistochemical analyses of MLH1 and MSH2 proteins. Among 44 MSI-H samples, sequence variations in BAT26 and BAT25 were detected in 44 (100%) and 43 (98%), respectively, while no sequence variation in the two markers was detected in 40 MSI-L or MSS samples. A total of 96 gastric cancers and their matched normal tissues were then analyzed for MSI-H using this method. Sequence variations in BAT26 and BAT25 were detected in nine (9.4%) samples. Seven of the nine cases were shown unstable at both BAT26 and BAT25; one each was unstable at BAT26 or BAT25. These results were confirmed by autoradiography analyses. Together, our results demonstrate high sensitivity and specificity of DHPLC in the analysis of sequence variations in BAT25 and BAT26 to determine MSI status. This simple, efficient, and high,throughput approach will facilitate analysis of MSI in large sample sets of any cancers. (C) 2003 Wiley-Liss, Inc.

语种英语
WOS记录号WOS:000186299100006
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被引频次:18[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/60466
专题北京大学临床肿瘤学院
作者单位1.Peking Univ, Beijing Canc Hosp, Dept Canc Epidemiol,Beijing Lab Mol Oncol, Sch Oncol,Beijing Inst Canc Res, Beijing, Peoples R China
2.Mayo Clin, Mayo Med Sch, Div Expt Pathol, Dept Pathol & Lab Med, Rochester, MN USA
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GB/T 7714
Pan, K,Liu, W,Lu, YY,et al. High throughput detection of microsatellite instability by denaturing high-performance liquid chromatography[J]. HUMAN MUTATION,2003,22(5):388-394.
APA Pan, K.,Liu, W.,Lu, YY.,Zhang, L.,Li, Z.,...&You, WC.(2003).High throughput detection of microsatellite instability by denaturing high-performance liquid chromatography.HUMAN MUTATION,22(5),388-394.
MLA Pan, K,et al."High throughput detection of microsatellite instability by denaturing high-performance liquid chromatography".HUMAN MUTATION 22.5(2003):388-394.
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