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学科主题: 临床医学
题名:
High throughput detection of microsatellite instability by denaturing high-performance liquid chromatography
作者: 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
发表日期: 2003-11-01
DOI: 10.1002/humu.10271
卷: 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
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/60466
Appears in Collections:北京大学临床肿瘤学院_期刊论文

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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

Recommended Citation:
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.
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