Mutation analysis of K-ras oncogenes in gastroenterologic cancers by the amplified created restriction sites method

S. Y. Lin, P. H. Chen, C. K. Wang, J. D. Liu, C. P. Siauw, Y. J. Chen, M. J. Yang -, M. H. Liu, T. C. Chen, J. G. Chang

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34 Citations (Scopus)

Abstract

A rapid, simple, and nonradioactive method for diagnosing point mutations of c-K-ras oncogenes in gastroenterologic cancers is described. This method involved the selective amplification of DNA fragments from cancer tissues of surgical specimens with specific oligonucleotide primers, followed by digestion with restriction enzymes that recognized artificially created or naturally occurring restriction sites. To detect codon 12 mutations, an artificial Msp I site was created by introducing a single nucleotide mismatch into the 5' mutagenesis primer. Using a similar approach, an Hae III site was created to detect codon 13 mutations. Bal I and MBo II sites were used to detect codon 61 mutations. A total of 61 gastroenterologic cancer cases were studied. Of 35 cases of colorectal cancer, 7 showed mutations: 6 at codon 12 and 1 at codon 13. In 1 of 2 cases of cholangiocellular carcinoma, point mutation at codon 12 was found. One case of duodenal cancer showed point mutation at codon 12. No mutations were found in the cases of hepatocellular carcinoma (4), gastric cancer (12), esophageal cancer (3), or pancreatic cancer (2).

Original languageEnglish
Pages (from-to)686-689
Number of pages4
JournalAmerican Journal of Clinical Pathology
Volume100
Issue number6
Publication statusPublished - 1993
Externally publishedYes

Keywords

  • Amplified created restriction site
  • Direct sequencing
  • Gastroenterologic cancers
  • Mutagenesis primer
  • Polymerase chain reaction
  • Ras oncogene
  • Restriction enzyme

ASJC Scopus subject areas

  • Medicine(all)

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  • Cite this

    Lin, S. Y., Chen, P. H., Wang, C. K., Liu, J. D., Siauw, C. P., Chen, Y. J., Yang -, M. J., Liu, M. H., Chen, T. C., & Chang, J. G. (1993). Mutation analysis of K-ras oncogenes in gastroenterologic cancers by the amplified created restriction sites method. American Journal of Clinical Pathology, 100(6), 686-689.