COMPARISON OF DIFFERENT METHODS OF MOLECULAR-GENETIC ANALYSIS OF SOMATIC MUTATIONS IN K-RAS GENE IN PATIENTS WITH COLORECTAL CANCER

Cover Page


Cite item

Full Text

Abstract

Two approaches to somatic point mutations in 12 and 13 codones of K-ras gene were analyzed: PCR/SSCP/AСRS/sequencing and allele-specific PCR in the real-life regimen (Russian set «KRAS-7M»). The comparison was carried out on 62 examples of genomic DNA extracted from frozen colon carcinomas, which underwent manual dissection. The results obtained in two attempts were consistent in 95,2% (N=59). Specificity and sensitivity of K-ras mutations detection using «KRAS-7M» set were 100 and 96,4% respectively, and 94,1 and 100% respectievly using PCR/SSCP/AСRS/ automatic sequencing. False positive results were absent when detecting with «KRAS-7M» and accounted for 2 cases (5,9%) when using PCR/SSCP/ AСRS/automatic sequencing. The only false negative response (3,6%) was obtained analyzing mutations using «KRAS-7M».

About the authors

F. A. Amosenko

Scientific centre of medical genetics, RAMS, Moscow
Blokhin Russian oncological scientific centre RAMS, Moscow

Author for correspondence.
Email: amossenko@med-gen.ru
кандидат биологических наук, старший научный сотрудник, ведущий научный сотрудник Медико-генетического научного центра РАМН, лаборатории ДНК-диагностики Адрес: 115478, Москва, ул. Москворечье, д. 1 Тел.: (965) 132-78-02, факс: (499) 324-07- 02 Russian Federation

I. V. Karpov

Laboratory of genetic diagnosis «BioLink», Novosibirsk

Email: biolink@inbox.ru
лаборант ООО «БиоЛинк» лаборатории генодиагностики Адрес: 630117, Новосибирск, ул. Академика Тимакова, д. 2 Тел./факс: (383) 334-86-14 Russian Federation

A. V. Polyakov

Scientific centre of medical genetics, RAMS, Moscow

Email: info@dnalab.ru
доктор биологических наук, профессор Медико-генетического научного центра РАМН, заведующий лабораторией ДНК-диагностики Адрес: 115478, Москва, ул. Москворечье, д. 1 Тел./факс: (495) 221-90-84 Russian Federation

S. P. Kovalenko

Institute of molecular biology and biophysics Siberian division of RAMS, Novosibirsk

Email: sp_kovalenko@yahoo.com
кандидат биологических наук, заведующий лабораторией генной инженерии Института молекулярной биологии и биофизики Сибирского Отделения РАМН Адрес: 630117, Новосибирск, ул. Академика Тимакова, д. 2 Тел/факс: (383) 333-47-10 Russian Federation

V. A. Shamanin

Laboratory of genetic diagnosis «BioLink», Novosibirsk

Email: va_shamanin@biolinklab.ru
кандидат биологических наук, ведущий научный сотрудник лаборатории гено- диагностики ООО «БиоЛинк» Адрес: 630117, Новосибирск, ул. Академика Тимакова, д. 2 Тел/факс: (383) 334-86-14 Russian Federation

L. N. Lyubchenko

Blokhin Russian oncological scientific centre RAMS, Moscow

Email: clingen@mail.ru
доктор медицинских наук, заведующая лабораторией клинической онкогенетики Российского онкологического научного центра им. Н.Н. Блохина РАМН Адрес: 115478, Москва, Каширское шоссе, д. 24 Тел.: (916) 623-23-66, факс: (499) 324-96-29 Russian Federation

References

  1. Marshall C.J. Small GTPases and cell cycle regulation. Biochem. Soc. Trans. 1999; 27 (4): 363−370.
  2. Chang F., Lee J.T., Navolanic P.M. et al. Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy. Leukemia. 2003; 17: 590−603.
  3. McCubrey J.A., Steelman L.S., Chappell W.H. et al. Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. Biochim. Biophys. Acta. 2007; 1773: 1263−1284.
  4. Bos J.L. К-ras oncogenes in human cancer: a review. Cancer Res. 1989; 49: 4682−4689.
  5. Minamoto T., Mai M., Ronai Z. K-ras mutation: early detection in molecular diagnosis and risk assessment of colorectal, pancreas, and lung cancers ― a review. Cancer Detect. Prev. 2000; 24: 1−12.
  6. Parkin D.M., Bray F., Ferlay J. et al. Global cancer statistics. 2002; CA. Cancer J. Clin. 2005; 55 (2): 74−108.
  7. Brink M., de Goeij A.F., Weijenberg M.P. et al. K-ras oncogene mutations in sporadic colorectal cancer in the Netherlands cohort study. Carcinogenesis. 2003; 24 (4): 703−710.
  8. Samowitz W.S., Curtin K., Schaffer D. et al. Relationship of Ki-ras mutations in colon cancers to tumor location, stage, and survival: a population-based study. Cancer Epidemial. Biomarkers Prev. 2000; 9 (11): 1193−1197.
  9. Slattery M.L., Curtin K., Anderson K. et al. Associations between dietary intake and Ki-ras mutations in colon tumors: a population-based study. Cancer Res. 2000; 60 (24): 6935−6941.
  10. Amosenko F.A., Korchagina E.L., Matveeva T.I. et al. Mutation analysis of K-ras protooncogene in colorectal adenocarcinomas and polyps in Russian patients. Russian J. Genetics. 2010; 46 (5): 617−624.
  11. Vogelstein B., Fearon E.R., Stanley S.R. et al. Genetic alterations during colorectal tumor development. N. Engl. J. Med. 1988; 319: 525−532.
  12. Lowy D.R., Willumsen B.M. Function and regulation of ras. Annu. Rev. Biochem. 1993; 62: 851−891.
  13. Al-Mulla F., Milner-White E.J., Going J.J. et al. Structural differences between valine-12 and aspartate-12 Ras proteins may modify carcinoma aggression. J. Pathol. 1999; 187: 433−438.
  14. Tuveson D.A., Shaw A.T., Willis N.A. et al. Endogenous oncogenic K-ras (G12D) stimulates proliferation and widespread neoplastic and developmental defects. Cancer Cell. 2004; 5: 375−387.
  15. Smakman N., Borel Rinkes I. H., Voest E. E. et al. Control of colorectal metastasis formation by K-ras. Biochim. Biophys. Acta. 2005; 1756: 103−114.
  16. Castagnola P., Giaretti W. Mutant KRAS, chromosomal instability and prognosis in colorectal cancer. Biochim. Biophys. Acta. 2005; 1756: 115−125.
  17. Pajkos G., Kiss I., Sandor J. et al. The prognostic value of the presence of mutations at the codons 12, 13, 61 of Ki-ras oncogene in colorectal cancer. Anticancer Res. 2000; 20: 1695−1701.
  18. Colin A., Smith G., Carey F. A. et al. The prognostic significance of K-ras, p53, and APC mutations in colorectal carcinoma. Gut. 2005; 54 (9): 1283−1286.
  19. Lievre A., Bachet J.B., Boige V. et al. KRAS mutations as an independent prognostic factor in patients with advanced colorectal cancer treated with cetuximab. J. Clin. Oncol. 2008; 26: 374−379.
  20. Amado R.G., Wolf M., Peeters M. et al. Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer. J. Clin. Oncol. 2008; 26: 1626−1634.
  21. Taniguchi K., Okami J., Kodama K. et al. Intratumor heterogeneity of epidermal growth factor receptor mutations in lung cancer and its correlation to the response to gefitinib. Cancer Sci. 2008; 99: 929−935.
  22. Fox J.C., England J., White P. et al. The detection of K-ras mutations in colorectal cancer using the amplification ― refractory mutation system. Br. J. Cancer. 1998; 77: 1267−1274.
  23. Poehlmann A., Kuester D., Meyer F. et al. K-ras mutation detection in colorectal cancer using the pyrosequencing technique. Pathol. Res. Pract. 2007; 203 (7): 489−497.
  24. Taback B., Bilchik A.J., Saha S. et al. Peptide nucleic acid clamp PCR: a novel K-ras mutation detection assay for colorectal cancer micrometastases in lymph nodes. Int. J. Cancer. 2004; 111 (3): 409−414.
  25. Lleonart M.E., Ramon Y., Cajal S. et al. Sensitive and specific detection of K-ras mutations in colon tumors by short oligonucleotide mass analysis. Nucl. Acids Res. 2004; 32: 53.
  26. Bjorheim J., Lystad S., Lindblom A. et al. Mutation analyses of KRAS exon 1 comparing three different techniques: temporal temperature gradient electrophoresis, constant denaturant capillary electrophoresis and allele specific polymerase reaction. Mutat. Res. 1998; 403: 103−112.
  27. Do H., Krypuy M., Mitchell P. et al. High resolution melting analysis for rapid and sensitive EGFR and KRAS mutation detection in formalin fixed paraffin embedded biopsies. BMC Cancer. 2008; 8: 142.
  28. Li J., Wang L., Mamon H. et al. Replacing PCR with COLD-PCR enriches variant DNA sequences and redefines the sensitivity of genetic testing. Nat. Med. 2008; 14 (5): 579−584.
  29. Ogino S., Kawasaki T., Brahmandam M. et al. Sensitive sequencing method for KRAS mutation detection by pyrosequencing. J. Mol. Diagn. 2005; 7 (3): 413−421.
  30. Ronaghi M., Uhlen M., Nyren P. A sequencing method based on real-time pyrophosphate. Science. 1998; 281 (5375): 363−365.
  31. Clayton S.J., Scott F.M., Walker J. et al. K-ras point mutation detection in lung cancer: comparison of two approaches to somatic mutation detection using ARMS allele-specific amplification. Clin. Chem. 2000; 46 (12): 1929−1938.
  32. Levi S., Urbano-Ispizua A., Gill R. et al. Multiple K-ras codon 12 mutations in cholangiocarcinomas demonstrated with a sensitive polymerase chain reaction technique. Cancer Res. 1991; 51: 3497−3502.
  33. Suzuki Y., Orita M., Shiraishi M. et al. Detection of ras gene mutations in human lung cancers by single-strand conformation polymorphism analysis of polymerase chain reaction products. Oncogene. 1990; 5: 1037−1043.
  34. Jean G.W., Shah S.R. Epidermal growth factor receptor monoclonal antibodies for the treatment of metastatic colorectal cancer. Pharmacotherapy. 2008; 28 (6): 742−754.
  35. Peeters M., Balfour J., Arnold D. Review article: panitumumab ― a fully human anti-EGFR monoclonal antibody for treatment of metastatic colorectal cancer. Aliment. Pharmacol. Ther. 2008; 28 (3): 269−281.
  36. Iqbal S., Lenz H.J. Integration of novel agents in the treatment of colorectal cancer. Cancer Chemother. Pharmacol. 2004; 54 (Suppl. 1): 32−39.
  37. de Castro-Carpeno J., Belda-Iniesta C., Casado Saenz E. et al. EGFR and colon cancer: a clinical view. Clin. Transl. Oncol. 2008; 10 (1): 6−13.
  38. Porebska I., Harlozinska A., Bojarowski T. Expression of the tyrosine kinase activity growth factor receptors (EGFR, ERB B2, ERB B3) in colorectal adenocarcinomas and adenomas. Tumour Biol. 2000; 21 (2): 105−115.
  39. Tigue C., Fitzner K., Alkhatib M. et al. The value of innovation: the economics of targeted drugs for cancer. Targeted Oncol. 2007; 2 (2): 113−119.
  40. Monzon F.A., Ogino S., Hammond E.H. et al. The role of KRAS mutation testing in the management of patients with metastatic colorectal cancer. Arch. Pathol. Lab. Med. 2009; 133: 1600−1606.

Supplementary files

Supplementary Files
Action
1. JATS XML
Download

Copyright (c) 2012 "Paediatrician" Publishers LLC



This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies