Detection of infrequent and multiple K-ras mutations in human tumors and tumor-adjacent tissues

Analytical Biochemistry
P KeohavongS D Finkelstein

Abstract

A sensitive method was developed and applied to examine the distribution of K-ras gene mutations in histologically differing areas of lung tissues obtained from lung cancer patients. This method, which combines polymerase chain reaction (PCR), mutation allele enrichment (MAE), and denaturing gradient gel electrophoresis (DGGE), allows detection of one K-ras mutant allele present in 10(4) to 10(5) wild-type alleles. It was applied to analyze mutations in codon 12 of the K-ras gene in 43 tissue sites microdissected from paraffin-embedded sections obtained from 8 archival cases of lung cancer, all previously shown to have codon 12 K-ras mutations by direct sequencing. In four cases, mutations were detected only in the tumor, while in the other four cases, the same mutations were also found in tissues adjacent to tumors, using the MAE + DGGE method. No mutations were detected among normal-appearing cells in areas distant from the tumors in any of the cases studied. These findings demonstrate that K-ras mutations can be detected at low frequencies in normal-appearing cells from tissues adjacent to the tumor in some lung cancer cases. In addition, this approach also allowed detection of multiple mutations in colorectal tissues obtain...Continue Reading

References

Jan 1, 1987·Annual Review of Biochemistry·M Barbacid
Dec 11, 1995·International Journal of Cancer. Journal International Du Cancer·M S YakubovskayaA C Trachtenberg

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Citations

Jul 29, 2005·The Journal of Molecular Diagnostics : JMD·Shuji OginoCharles S Fuchs
Sep 1, 2010·Expert Opinion on Medical Diagnostics·Gernot KriegshäuserRobert Zeillinger
May 31, 2003·Biomarkers : Biochemical Indicators of Exposure, Response, and Susceptibility to Chemicals·Li-Fang ZhangPhouthone Keohavong
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May 18, 2016·Molecular Carcinogenesis·Weimin GaoPhouthone Keohavong
May 20, 2000·Annals of the New York Academy of Sciences·C D GockeD B Evans

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