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Molecular Medicine

, Volume 18, Issue 12, pp 1519–1526 | Cite as

Low Percentage of KRAS Mutations Revealed by Locked Nucleic Acid Polymerase Chain Reaction: Implications for Treatment of Metastatic Colorectal Cancer

  • Mariella Dono
  • Carlotta Massucco
  • Silvana Chiara
  • Claudia Sonaglio
  • Marco Mora
  • Anna Truini
  • Giannamaria Cerruti
  • Gabriele Zoppoli
  • Alberto Ballestrero
  • Mauro Truini
  • Manlio Ferrarini
  • Simona Zupo
Research Article

Abstract

Metastatic colorectal cancer (mCRC) is frequently characterized by the presence of mutations of the KRAS oncogene, which are generally associated with a poor response to treatment with anti-epidermal growth factor receptor (anti-EGFR) monoclonal antibodies. With the methods currently used, a case is classified as KRAS-mutated when approximately 20% of the cells bear an activating KRAS mutation. These considerations raise the question of whether cells with a mutated KRAS can be found in mCRC cases classified as KRAS wild-type when more sensitive methods are used. In addition, the issue arises of whether these mCRC cases with low proportion of KRAS-mutated cells could account at least in part for the therapeutic failure of anti-EGFR therapies that occur in 40–60% of cases classified as KRAS wild type. In this study, we compared the classical assays with a very sensitive test, a locked nucleic acid (LNA) polymerase chain reaction (PCR), capable of detecting KRAS-mutated alleles at extremely low frequency (detection sensitivity limit 0.25% mutated DNA/wild-type DNA). By analyzing a cohort of 213 mCRC patients for KRAS mutations, we found a 20.6% discordance between the sequencing/TheraScreen methods and the LNA-PCR. Indeed, 44 mCRC patients initially considered KRAS wild type were reclassified as KRAS mutated by using the LNA-PCR test. These patients were more numerous among individuals displaying a clinical failure to anti-EGFR therapies. Failure to respond to these biological treatments occurred even in the absence of mutations in other EGFR pathway components such as BRAF.

Notes

Acknowledgments

This research was supported by Regione Liguria: “Diagnostica Oncologica Avanzata” and “Introducing new laboratory tests in clinical practice and oncological networks.”

C Massucco is a postdoc fellow supported by Associazione Italiana per la Ricerca sul Cancro (AIRC). The authors thank Floriana Pensa for collecting clinical data.

Supplementary material

10020_2012_18121519_MOESM1_ESM.pdf (337 kb)
Low Percentage of KRAS Mutations Revealed by Locked Nucleic Acid Polymerase Chain Reaction: Implications for Treatment of Metastatic Colorectal Cancer

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© The Author(s) 2012

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Authors and Affiliations

  • Mariella Dono
    • 1
    • 2
  • Carlotta Massucco
    • 1
  • Silvana Chiara
    • 3
  • Claudia Sonaglio
    • 3
  • Marco Mora
    • 2
  • Anna Truini
    • 1
  • Giannamaria Cerruti
    • 1
  • Gabriele Zoppoli
    • 4
  • Alberto Ballestrero
    • 4
  • Mauro Truini
    • 2
  • Manlio Ferrarini
    • 5
  • Simona Zupo
    • 1
  1. 1.Molecular Diagnostic DivisionIstituto di Ricovero e Cura a Carattere Scientitifico Azienda Ospedaliera Universitaria San Martino-Istituto Nazionale per la Ricerca sul Cancro (IRCCS AOU San Martino-IST)GenovaItaly
  2. 2.Department of PathologyIstituto di Ricovero e Cura a Carattere Scientitifico Azienda Ospedaliera Universitaria San Martino-Istituto Nazionale per la Ricerca sul Cancro (IRCCS AOU San Martino-IST)GenovaItaly
  3. 3.Medical Oncology DivisionIstituto di Ricovero e Cura a Carattere Scientitifico Azienda Ospedaliera Universitaria San Martino-Istituto Nazionale per la Ricerca sul Cancro (IRCCS AOU San Martino-IST)GenovaItaly
  4. 4.Division of Semeiotics and Medical Methodology, Department of Internal MedicineIstituto di Ricovero e Cura a Carattere Scientitifico Azienda Ospedaliera Universitaria San Martino-Istituto Nazionale per la Ricerca sul Cancro (IRCCS AOU San Martino-IST)GenovaItaly
  5. 5.Direzione ScientificaIstituto di Ricovero e Cura a Carattere Scientitifico Azienda Ospedaliera Universitaria San Martino-Istituto Nazionale per la Ricerca sul Cancro (IRCCS AOU San Martino-IST)GenovaItaly

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