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Detection of ras gene mutations in human lung cancer: comparison of two screening assays based on the polymerase chain reaction.

作者信息

Husgafvel-Pursiainen K, Ridanpää M, Hackman P, Anttila S, Karjalainen A, Onfelt A, Børresen A L, Vainio H

机构信息

Institute of Occupational Health, Helsinki, Finland.

出版信息

Environ Health Perspect. 1992 Nov;98:183-5. doi: 10.1289/ehp.9298183.

DOI:10.1289/ehp.9298183
PMID:1486847
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1519635/
Abstract

We studied the prevalence of point mutations in ras oncogenes (K-ras and N-ras) in DNA from white blood cells and tumor tissue from 36 untreated patients with non-small-cell lung cancer, all of whom were smokers or ex-smokers. We observed somatic K-ras mutations in one-third of the lung carcinomas studied but no N-ras mutation. K-ras codon 12 mutations were found more frequently in adenocarcinomas than in the other histopathological subtypes studied. More than 60% (10/16) of the lung adenocarcinomas had a codon 12 mutation, most of which were G to T transversions. No mutations was found in white blood cell DNA. Two polymerase chain reaction screening methods, oligonucleotide hybridization and denaturing gradient gel electrophoresis (DGGE), were used to detect the mutations. The oligonucleotide method appears to be more sensitive than DGGE, but DGGE proved to be a reliable nonradioactive method for rapid screening of point mutations in genes relevant to carcinogenesis.

摘要
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/44fe/1519635/ae9ce5431df8/envhper00385-0180-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/44fe/1519635/e65b92b2549c/envhper00385-0180-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/44fe/1519635/ae9ce5431df8/envhper00385-0180-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/44fe/1519635/e65b92b2549c/envhper00385-0180-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/44fe/1519635/ae9ce5431df8/envhper00385-0180-b.jpg

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本文引用的文献

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DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: correspondence with melting theory.通过单碱基对替换而不同的DNA片段在变性梯度凝胶中被分离:与解链理论的对应关系。
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Attachment of a 40-base-pair G + C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes.通过聚合酶链反应将一段40个碱基对的富含G + C的序列(GC夹)连接到基因组DNA片段上,可提高对单碱基变化的检测。
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ras oncogenes in human cancer: a review.人类癌症中的Ras癌基因:综述
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8
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.使用热稳定DNA聚合酶进行引物引导的DNA酶促扩增。
Science. 1988 Jan 29;239(4839):487-91. doi: 10.1126/science.2448875.
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Cancer. A deadly inheritance.
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N Engl J Med. 1990 Aug 30;323(9):561-5. doi: 10.1056/NEJM199008303230902.