Detection of K-Ras mutations in tumour samples of patients with non-small cell lung cancer using PNA-mediated PCR clamping.
Fiche publication
Date publication
mars 2009
Auteurs
Membres identifiés du Cancéropôle Est :
Dr BEAU-FALLER Michèle, Dr GUERIN Eric, Pr OUDET Pierre
Tous les auteurs :
Beau-Faller M, Legrain M, Voegeli AC, Guerin E, Lavaux T, Ruppert AM, Neuville A, Massard G, Wihlm JM, Quoix E, Oudet P, Gaub MP
Lien Pubmed
Résumé
Non-small cell lung cancers (NSCLC), in particular adenocarcinoma, are often mixed with normal cells. Therefore, low sensitivity of direct sequencing used for K-Ras mutation analysis could be inadequate in some cases. Our study focused on the possibility to increase the detection of K-Ras mutations in cases of low tumour cellularity. Besides direct sequencing, we used wild-type hybridisation probes and peptide-nucleic-acid (PNA)-mediated PCR clamping to detect mutations at codons 12 and 13, in 114 routine consecutive NSCLC frozen surgical tumours untreated by targeted drugs. The sensitivity of the analysis without or with PNA was 10 and 1% of tumour DNA, respectively. Direct sequencing revealed K-Ras mutations in 11 out of 114 tumours (10%). Using PNA-mediated PCR clamping, 10 additional cases of K-Ras mutations were detected (21 out of 114, 18%, P
Référence
Br J Cancer. 2009 Mar 24;100(6):985-92.