open access

Vol 4, No 6 (2008)
Research paper
Published online: 2008-12-16
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Validation of selected molecular methods for the mutations determination in codons 12 and 13 of K-RAS gene in five Polish oncological research centers

Andrzej Tysarowski, Anna Fabisiewicz, Iwona Kolasa, Jolanta Kupryjańczyk, Dorota Ścieglińska, Marek Rusin, Zdzisław Krawczyk, Agnieszka Woźniak, Lucyna Morzuch, Janusz Limon, Oksana Kowalczuk, Lech Chyczewski, Piotr Wójcik, Jerzy Stachura, Karolina Wieruszewska-Kowalczyk, Monika Pokrzepa, Włodzimierz P. Olszewski, Marek P. Nowacki, Janusz A. Siedlecki
Onkol. Prak. Klin 2008;4(6):232-244.

open access

Vol 4, No 6 (2008)
ORIGINAL ARTICLES
Published online: 2008-12-16

Abstract

Metastatic colorectal cancer patients will benefit from treatment with panitumumab only when they don't have mutation in K-RAS gene. Therefore, estimation of mutational status of K-RAS is necessary for the selection of patients, who should be treated with panitumumab.
The aim of this study was to evolve a standard method of estimation of K-RAS mutational status in the material isolated from paraffin blocs. The second aim was the validation of selected molecular methods of K-RAS mutation evaluation in five Polish oncological centers where mCRC patients are treated. Four methods were evaluated: SSCP, DHPLC, RFLP/PCR and direct sequencing.
We found that all groups in five selected oncological centers, who took part in the validation process, were well prepared for molecular diagnosis of K-RAS mutational status. The following recommendations for diagnostic laboratories were approved: 1. At least 70% of cancer cells should be present in a tissue for DNA isolation; 2. The method of DNA isolation should be standardized, the most appropriate is usage of DNA isolation kits; 3. In case of equivocal results two independent molecular methods should be employed, one of them should be direct sequencing.

Abstract

Metastatic colorectal cancer patients will benefit from treatment with panitumumab only when they don't have mutation in K-RAS gene. Therefore, estimation of mutational status of K-RAS is necessary for the selection of patients, who should be treated with panitumumab.
The aim of this study was to evolve a standard method of estimation of K-RAS mutational status in the material isolated from paraffin blocs. The second aim was the validation of selected molecular methods of K-RAS mutation evaluation in five Polish oncological centers where mCRC patients are treated. Four methods were evaluated: SSCP, DHPLC, RFLP/PCR and direct sequencing.
We found that all groups in five selected oncological centers, who took part in the validation process, were well prepared for molecular diagnosis of K-RAS mutational status. The following recommendations for diagnostic laboratories were approved: 1. At least 70% of cancer cells should be present in a tissue for DNA isolation; 2. The method of DNA isolation should be standardized, the most appropriate is usage of DNA isolation kits; 3. In case of equivocal results two independent molecular methods should be employed, one of them should be direct sequencing.
Get Citation

Keywords

metastatic colorectal cancer; K-RAS mutation; K-RAS mutational status; panitumumab

About this article
Title

Validation of selected molecular methods for the mutations determination in codons 12 and 13 of K-RAS gene in five Polish oncological research centers

Journal

Oncology in Clinical Practice

Issue

Vol 4, No 6 (2008)

Article type

Research paper

Pages

232-244

Published online

2008-12-16

Bibliographic record

Onkol. Prak. Klin 2008;4(6):232-244.

Keywords

metastatic colorectal cancer
K-RAS mutation
K-RAS mutational status
panitumumab

Authors

Andrzej Tysarowski
Anna Fabisiewicz
Iwona Kolasa
Jolanta Kupryjańczyk
Dorota Ścieglińska
Marek Rusin
Zdzisław Krawczyk
Agnieszka Woźniak
Lucyna Morzuch
Janusz Limon
Oksana Kowalczuk
Lech Chyczewski
Piotr Wójcik
Jerzy Stachura
Karolina Wieruszewska-Kowalczyk
Monika Pokrzepa
Włodzimierz P. Olszewski
Marek P. Nowacki
Janusz A. Siedlecki

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