Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1998 Apr;77(8):1267–1274. doi: 10.1038/bjc.1998.212

The detection of K-ras mutations in colorectal cancer using the amplification-refractory mutation system.

J C Fox 1, J England 1, P White 1, G Ellison 1, K Callaghan 1, N R Charlesworth 1, J Hehir 1, T L McCarthy 1, J Smith-Ravin 1, I C Talbot 1, D Snary 1, J M Northover 1, C R Newton 1, S Little 1
PMCID: PMC2150152  PMID: 9579832

Abstract

A total of 301 colorectal carcinoma (CRC) archival samples were analysed using the amplification-refractory mutation system (ARMS). Each sample was examined to determine the mutation status of codons 12 and 13 of the K-ras oncogene. The results from direct DNA sequence analysis carried out on 30 of the samples differed from the ARMS result in almost 50% of the cases as a result of the relative excess of wild-type to mutated DNA sequences. To assess the validity of the ARMS data, the polymerase chain reaction (PCR) was used to generate an amplicon from K-ras exon 1 from 23 of the samples. The PCR amplicons were cloned and sequenced, and the DNA sequence analysis of the cloned material was in agreement with the ARMS results in all but one case. This case represented a tumour that exhibited a five-nucleotide reversed inversion. The cloned sequence data confirm the sensitivity and specificity of the individual ARMS reactions and that it is possible in certain cases to detect additional, more complex, sequence variations.

Full text

PDF
1267

Images in this article

1269Figure 2
on p.1269
1270Figure 3
on p.1270
1270Figure 4
on p.1270

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bell S. M., Scott N., Cross D., Sagar P., Lewis F. A., Blair G. E., Taylor G. R., Dixon M. F., Quirke P. Prognostic value of p53 overexpression and c-Ki-ras gene mutations in colorectal cancer. Gastroenterology. 1993 Jan;104(1):57–64. doi: 10.1016/0016-5085(93)90835-z. [DOI] [PubMed] [Google Scholar]
  2. Bos J. L., Fearon E. R., Hamilton S. R., Verlaan-de Vries M., van Boom J. H., van der Eb A. J., Vogelstein B. Prevalence of ras gene mutations in human colorectal cancers. 1987 May 28-Jun 3Nature. 327(6120):293–297. doi: 10.1038/327293a0. [DOI] [PubMed] [Google Scholar]
  3. Bos J. L. ras oncogenes in human cancer: a review. Cancer Res. 1989 Sep 1;49(17):4682–4689. [PubMed] [Google Scholar]
  4. Breivik J., Meling G. I., Spurkland A., Rognum T. O., Gaudernack G. K-ras mutation in colorectal cancer: relations to patient age, sex and tumour location. Br J Cancer. 1994 Feb;69(2):367–371. doi: 10.1038/bjc.1994.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burmer G. C., Loeb L. A. Mutations in the KRAS2 oncogene during progressive stages of human colon carcinoma. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2403–2407. doi: 10.1073/pnas.86.7.2403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Capella G., Cronauer-Mitra S., Pienado M. A., Perucho M. Frequency and spectrum of mutations at codons 12 and 13 of the c-K-ras gene in human tumors. Environ Health Perspect. 1991 Jun;93:125–131. doi: 10.1289/ehp.9193125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carpenter K. M., Durrant L. G., Morgan K., Bennett D., Hardcastle J. D., Kalsheker N. A. Greater frequency of K-ras Val-12 mutation in colorectal cancer as detected with sensitive methods. Clin Chem. 1996 Jun;42(6 Pt 1):904–909. [PubMed] [Google Scholar]
  8. Cha R. S., Zarbl H., Keohavong P., Thilly W. G. Mismatch amplification mutation assay (MAMA): application to the c-H-ras gene. PCR Methods Appl. 1992 Aug;2(1):14–20. doi: 10.1101/gr.2.1.14. [DOI] [PubMed] [Google Scholar]
  9. Chen J., Viola M. V. A method to detect ras point mutations in small subpopulations of cells. Anal Biochem. 1991 May 15;195(1):51–56. doi: 10.1016/0003-2697(91)90293-3. [DOI] [PubMed] [Google Scholar]
  10. Chou Q., Russell M., Birch D. E., Raymond J., Bloch W. Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res. 1992 Apr 11;20(7):1717–1723. doi: 10.1093/nar/20.7.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. D'Aquila R. T., Bechtel L. J., Videler J. A., Eron J. J., Gorczyca P., Kaplan J. C. Maximizing sensitivity and specificity of PCR by pre-amplification heating. Nucleic Acids Res. 1991 Jul 11;19(13):3749–3749. doi: 10.1093/nar/19.13.3749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Delattre O., Olschwang S., Law D. J., Melot T., Remvikos Y., Salmon R. J., Sastre X., Validire P., Feinberg A. P., Thomas G. Multiple genetic alterations in distal and proximal colorectal cancer. Lancet. 1989 Aug 12;2(8659):353–356. doi: 10.1016/s0140-6736(89)90537-0. [DOI] [PubMed] [Google Scholar]
  13. Dix B. R., Robbins P., Soong R., Jenner D., House A. K., Iacopetta B. J. The common molecular genetic alterations in Dukes' B and C colorectal carcinomas are not short-term prognostic indicators of survival. Int J Cancer. 1994 Dec 15;59(6):747–751. doi: 10.1002/ijc.2910590606. [DOI] [PubMed] [Google Scholar]
  14. Ehlen T., Dubeau L. Detection of ras point mutations by polymerase chain reaction using mutation-specific, inosine-containing oligonucleotide primers. Biochem Biophys Res Commun. 1989 Apr 28;160(2):441–447. doi: 10.1016/0006-291x(89)92452-2. [DOI] [PubMed] [Google Scholar]
  15. Fearon E. R., Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990 Jun 1;61(5):759–767. doi: 10.1016/0092-8674(90)90186-i. [DOI] [PubMed] [Google Scholar]
  16. Ferrie R. M., Schwarz M. J., Robertson N. H., Vaudin S., Super M., Malone G., Little S. Development, multiplexing, and application of ARMS tests for common mutations in the CFTR gene. Am J Hum Genet. 1992 Aug;51(2):251–262. [PMC free article] [PubMed] [Google Scholar]
  17. Finkelstein S. D., Sayegh R., Bakker A., Swalsky P. Determination of tumor aggressiveness in colorectal cancer by K-ras-2 analysis. Arch Surg. 1993 May;128(5):526–532. doi: 10.1001/archsurg.1993.01420170056008. [DOI] [PubMed] [Google Scholar]
  18. Finkelstein S. D., Sayegh R., Christensen S., Swalsky P. A. Genotypic classification of colorectal adenocarcinoma. Biologic behavior correlates with K-ras-2 mutation type. Cancer. 1993 Jun 15;71(12):3827–3838. doi: 10.1002/1097-0142(19930615)71:12<3827::aid-cncr2820711207>3.0.co;2-n. [DOI] [PubMed] [Google Scholar]
  19. Forrester K., Almoguera C., Han K., Grizzle W. E., Perucho M. Detection of high incidence of K-ras oncogenes during human colon tumorigenesis. 1987 May 28-Jun 3Nature. 327(6120):298–303. doi: 10.1038/327298a0. [DOI] [PubMed] [Google Scholar]
  20. Giaretti W., Pujic N., Rapallo A., Nigro S., Di Vinci A., Geido E., Risio M. K-ras-2 G-C and G-T transversions correlate with DNA aneuploidy in colorectal adenomas. Gastroenterology. 1995 Apr;108(4):1040–1047. doi: 10.1016/0016-5085(95)90201-5. [DOI] [PubMed] [Google Scholar]
  21. Goldgar D. E., Easton D. F., Cannon-Albright L. A., Skolnick M. H. Systematic population-based assessment of cancer risk in first-degree relatives of cancer probands. J Natl Cancer Inst. 1994 Nov 2;86(21):1600–1608. doi: 10.1093/jnci/86.21.1600. [DOI] [PubMed] [Google Scholar]
  22. Green P. M., Bentley D. R., Mibashan R. S., Nilsson I. M., Giannelli F. Molecular pathology of haemophilia B. EMBO J. 1989 Apr;8(4):1067–1072. doi: 10.1002/j.1460-2075.1989.tb03474.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hasegawa Y., Takeda S., Ichii S., Koizumi K., Maruyama M., Fujii A., Ohta H., Nakajima T., Okuda M., Baba S. Detection of K-ras mutations in DNAs isolated from feces of patients with colorectal tumors by mutant-allele-specific amplification (MASA). Oncogene. 1995 Apr 6;10(7):1441–1445. [PubMed] [Google Scholar]
  24. Hayashi N., Ito I., Yanagisawa A., Kato Y., Nakamori S., Imaoka S., Watanabe H., Ogawa M., Nakamura Y. Genetic diagnosis of lymph-node metastasis in colorectal cancer. Lancet. 1995 May 20;345(8960):1257–1259. doi: 10.1016/s0140-6736(95)90922-2. [DOI] [PubMed] [Google Scholar]
  25. Houlston R. S., Collins A., Slack J., Morton N. E. Dominant genes for colorectal cancer are not rare. Ann Hum Genet. 1992 May;56(Pt 2):99–103. doi: 10.1111/j.1469-1809.1992.tb01136.x. [DOI] [PubMed] [Google Scholar]
  26. Jones P. A., Buckley J. D. The role of DNA methylation in cancer. Adv Cancer Res. 1990;54:1–23. doi: 10.1016/s0065-230x(08)60806-4. [DOI] [PubMed] [Google Scholar]
  27. Kern S. E., Fearon E. R., Tersmette K. W., Enterline J. P., Leppert M., Nakamura Y., White R., Vogelstein B., Hamilton S. R. Clinical and pathological associations with allelic loss in colorectal carcinoma [corrected]. JAMA. 1989 Jun 2;261(21):3099–3103. doi: 10.1001/jama.261.21.3099. [DOI] [PubMed] [Google Scholar]
  28. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  29. Laird P. W., Jackson-Grusby L., Fazeli A., Dickinson S. L., Jung W. E., Li E., Weinberg R. A., Jaenisch R. Suppression of intestinal neoplasia by DNA hypomethylation. Cell. 1995 Apr 21;81(2):197–205. doi: 10.1016/0092-8674(95)90329-1. [DOI] [PubMed] [Google Scholar]
  30. Laurent-Puig P., Olschwang S., Delattre O., Remvikos Y., Asselain B., Melot T., Validire P., Muleris M., Girodet J., Salmon R. J. Survival and acquired genetic alterations in colorectal cancer. Gastroenterology. 1992 Apr;102(4 Pt 1):1136–1141. [PubMed] [Google Scholar]
  31. Levi S., Urbano-Ispizua A., Gill R., Thomas D. M., Gilbertson J., Foster C., Marshall C. J. Multiple K-ras codon 12 mutations in cholangiocarcinomas demonstrated with a sensitive polymerase chain reaction technique. Cancer Res. 1991 Jul 1;51(13):3497–3502. [PubMed] [Google Scholar]
  32. Lewis C. M., Neuhausen S. L., Daley D., Black F. J., Swensen J., Burt R. W., Cannon-Albright L. A., Skolnick M. H. Genetic heterogeneity and unmapped genes for colorectal cancer. Cancer Res. 1996 Mar 15;56(6):1382–1388. [PubMed] [Google Scholar]
  33. MACKLIN M. T. Inheritance of cancer of the stomach and large intestine in man. J Natl Cancer Inst. 1960 Mar;24:551–571. doi: 10.1093/jnci/24.3.551. [DOI] [PubMed] [Google Scholar]
  34. Magewu A. N., Jones P. A. Ubiquitous and tenacious methylation of the CpG site in codon 248 of the p53 gene may explain its frequent appearance as a mutational hot spot in human cancer. Mol Cell Biol. 1994 Jun;14(6):4225–4232. doi: 10.1128/mcb.14.6.4225-4232.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McLellan E. A., Owen R. A., Stepniewska K. A., Sheffield J. P., Lemoine N. R. High frequency of K-ras mutations in sporadic colorectal adenomas. Gut. 1993 Mar;34(3):392–396. doi: 10.1136/gut.34.3.392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Moerkerk P., Arends J. W., van Driel M., de Bruïne A., de Goeij A., ten Kate J. Type and number of Ki-ras point mutations relate to stage of human colorectal cancer. Cancer Res. 1994 Jul 1;54(13):3376–3378. [PubMed] [Google Scholar]
  37. Morrin M., Kelly M., Barrett N., Delaney P. Mutations of Ki-ras and p53 genes in colorectal cancer and their prognostic significance. Gut. 1994 Nov;35(11):1627–1631. doi: 10.1136/gut.35.11.1627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Newton C. R., Graham A., Heptinstall L. E., Powell S. J., Summers C., Kalsheker N., Smith J. C., Markham A. F. Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Res. 1989 Apr 11;17(7):2503–2516. doi: 10.1093/nar/17.7.2503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Newton C. R., Kalsheker N., Graham A., Powell S., Gammack A., Riley J., Markham A. F. Diagnosis of alpha 1-antitrypsin deficiency by enzymatic amplification of human genomic DNA and direct sequencing of polymerase chain reaction products. Nucleic Acids Res. 1988 Sep 12;16(17):8233–8243. doi: 10.1093/nar/16.17.8233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Offerhaus G. J., De Feyter E. P., Cornelisse C. J., Tersmette K. W., Floyd J., Kern S. E., Vogelstein B., Hamilton S. R. The relationship of DNA aneuploidy to molecular genetic alterations in colorectal carcinoma. Gastroenterology. 1992 May;102(5):1612–1619. doi: 10.1016/0016-5085(92)91721-f. [DOI] [PubMed] [Google Scholar]
  41. Oudejans J. J., Slebos R. J., Zoetmulder F. A., Mooi W. J., Rodenhuis S. Differential activation of ras genes by point mutation in human colon cancer with metastases to either lung or liver. Int J Cancer. 1991 Dec 2;49(6):875–879. doi: 10.1002/ijc.2910490613. [DOI] [PubMed] [Google Scholar]
  42. Peltomäki P., Aaltonen L. A., Sistonen P., Pylkkänen L., Mecklin J. P., Järvinen H., Green J. S., Jass J. R., Weber J. L., Leach F. S. Genetic mapping of a locus predisposing to human colorectal cancer. Science. 1993 May 7;260(5109):810–812. doi: 10.1126/science.8484120. [DOI] [PubMed] [Google Scholar]
  43. Petersen G. M. Genetic epidemiology of colorectal cancer. Eur J Cancer. 1995 Jul-Aug;31A(7-8):1047–1050. doi: 10.1016/0959-8049(95)00156-d. [DOI] [PubMed] [Google Scholar]
  44. Ranaldi R., Gioacchini A. M., Manzin A., Clementi M., Paolucci S., Bearzi I. Adenoma-carcinoma sequence of colorectum. Prevalence of K-ras gene mutation in adenomas with increasing degree of dysplasia and aneuploidy. Diagn Mol Pathol. 1995 Sep;4(3):198–202. [PubMed] [Google Scholar]
  45. Redston M. S., Kern S. E. Klenow co-sequencing: a method for eliminating "stops". Biotechniques. 1994 Aug;17(2):286–288. [PubMed] [Google Scholar]
  46. Riley J., Jenner D., Smith J. C., Markham A. F. Rapid determination of DNA concentration in multiple samples. Nucleic Acids Res. 1989 Oct 25;17(20):8383–8383. doi: 10.1093/nar/17.20.8383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Riordan J. R., Rommens J. M., Kerem B., Alon N., Rozmahel R., Grzelczak Z., Zielenski J., Lok S., Plavsic N., Chou J. L. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science. 1989 Sep 8;245(4922):1066–1073. doi: 10.1126/science.2475911. [DOI] [PubMed] [Google Scholar]
  48. Sharrard R. M., Royds J. A., Rogers S., Shorthouse A. J. Patterns of methylation of the c-myc gene in human colorectal cancer progression. Br J Cancer. 1992 May;65(5):667–672. doi: 10.1038/bjc.1992.142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Sidransky D., Tokino T., Hamilton S. R., Kinzler K. W., Levin B., Frost P., Vogelstein B. Identification of ras oncogene mutations in the stool of patients with curable colorectal tumors. Science. 1992 Apr 3;256(5053):102–105. doi: 10.1126/science.1566048. [DOI] [PubMed] [Google Scholar]
  50. Span M., Moerkerk P. T., De Goeij A. F., Arends J. W. A detailed analysis of K-ras point mutations in relation to tumor progression and survival in colorectal cancer patients. Int J Cancer. 1996 Jun 21;69(3):241–245. doi: 10.1002/(SICI)1097-0215(19960621)69:3<241::AID-IJC15>3.0.CO;2-A. [DOI] [PubMed] [Google Scholar]
  51. Stork P., Loda M., Bosari S., Wiley B., Poppenhusen K., Wolfe H. Detection of K-ras mutations in pancreatic and hepatic neoplasms by non-isotopic mismatched polymerase chain reaction. Oncogene. 1991 May;6(5):857–862. [PubMed] [Google Scholar]
  52. Tanaka M., Omura K., Watanabe Y., Oda Y., Nakanishi I. Prognostic factors of colorectal cancer: K-ras mutation, overexpression of the p53 protein, and cell proliferative activity. J Surg Oncol. 1994 Sep;57(1):57–64. doi: 10.1002/jso.2930570115. [DOI] [PubMed] [Google Scholar]
  53. Urosević N., Krtolica K., Skaro-Milić A., Knezević-Usaj S., Dujić A. Prevalence of G-to-T transversions among K-ras oncogene mutations in human colorectal tumors in Yugoslavia. Int J Cancer. 1993 May 8;54(2):249–254. doi: 10.1002/ijc.2910540215. [DOI] [PubMed] [Google Scholar]
  54. Villa E., Dugani A., Rebecchi A. M., Vignoli A., Grottola A., Buttafoco P., Losi L., Perini M., Trande P., Merighi A. Identification of subjects at risk for colorectal carcinoma through a test based on K-ras determination in the stool. Gastroenterology. 1996 May;110(5):1346–1353. doi: 10.1053/gast.1996.v110.pm8613038. [DOI] [PubMed] [Google Scholar]
  55. Vogelstein B., Fearon E. R., Hamilton S. R., Kern S. E., Preisinger A. C., Leppert M., Nakamura Y., White R., Smits A. M., Bos J. L. Genetic alterations during colorectal-tumor development. N Engl J Med. 1988 Sep 1;319(9):525–532. doi: 10.1056/NEJM198809013190901. [DOI] [PubMed] [Google Scholar]
  56. Vogelstein B., Fearon E. R., Kern S. E., Hamilton S. R., Preisinger A. C., Nakamura Y., White R. Allelotype of colorectal carcinomas. Science. 1989 Apr 14;244(4901):207–211. doi: 10.1126/science.2565047. [DOI] [PubMed] [Google Scholar]
  57. WOOLF C. M. A genetic study of carcinoma of the large intestine. Am J Hum Genet. 1958 Mar;10(1):42–47. [PMC free article] [PubMed] [Google Scholar]
  58. Yamada H., Yoshida T., Sakamoto H., Terada M., Sugimura T. Establishment of a human pancreatic adenocarcinoma cell line (PSN-1) with amplifications of both c-myc and activated c-Ki-ras by a point mutation. Biochem Biophys Res Commun. 1986 Oct 15;140(1):167–173. doi: 10.1016/0006-291x(86)91072-7. [DOI] [PubMed] [Google Scholar]
  59. Young G., Levin B. Report of UICC colorectal cancer screening workshop. Meeting held in Genoa, Italy, September 29-October 1, 1995. Int J Cancer. 1996 Mar 1;65(5):567–568. doi: 10.1002/(SICI)1097-0215(19960301)65:5<567::AID-IJC2>3.0.CO;2-0. [DOI] [PubMed] [Google Scholar]
  60. Zhao L. P., Le Marchand L. An analytical method for assessing patterns of familial aggregation in case-control studies. Genet Epidemiol. 1992;9(2):141–154. doi: 10.1002/gepi.1370090206. [DOI] [PubMed] [Google Scholar]
  61. el-Deiry W. S., Nelkin B. D., Celano P., Yen R. W., Falco J. P., Hamilton S. R., Baylin S. B. High expression of the DNA methyltransferase gene characterizes human neoplastic cells and progression stages of colon cancer. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3470–3474. doi: 10.1073/pnas.88.8.3470. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES