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. 2013 Aug;139(8):1327-35.
doi: 10.1007/s00432-013-1444-y. Epub 2013 May 5.

EGFR and KRAS mutational profiling in fresh non-small cell lung cancer (NSCLC) cells

Giulia M Stella  1 , Roberta Scabini, Simona Inghilleri, Francesca Cemmi, Simona Corso, Ernesto Pozzi, Patrizia Morbini, Adele Valentini, Roberto Dore, Simona Ferrari, Maurizio Luisetti, Michele Zorzetto
Affiliations

EGFR and KRAS mutational profiling in fresh non-small cell lung cancer (NSCLC) cells

Giulia M Stella et al. J Cancer Res Clin Oncol. 2013 Aug.

Abstract

Purpose: Knowledge of tumor mutational status has become a priority for effective NSCLC-tailored treatment. NSCLC diagnosis is more often reached through biopsy; thus, there is a clear need to implement for routine tumor molecular profiling on small cytological samples. This work aims to screen and compare the EGFR and KRAS mutational prevalence in fresh tumor cells and in corresponding routinely processed samples derived from trans-thoracic fine-needle aspiration. The latter currently represents the most appropriate diagnostic procedure in case of peripheral lesions, such as adenocarcinomas, which account for almost 40% of all NSCLCs and for the highest EGFR mutational rates.

Methods: Two hundred and forty-four patients carrying peripheral lung masses underwent CT-guided aspiration. The obtained material was split, and a part was addressed to conventional histopathological analysis while the remaining one was stored at -20 °C. In case of confirmation of adenocarcinoma, tumor genomic DNA was extracted from both fresh and fixed material, and EGFR and KRAS sequencing was performed.

Results: We identified 136 adenocarcinomas; from 134, we could recover enough material for the study. A full match was demonstrated between EGFR/KRAS mutational prevalences through the two approaches tested. We found EGFR mutations in 13 patients (9.7%); 7 were females and 11 never or former smokers. KRAS mutations occurred in 20 (14.9%) patients. EGFR and KRAS mutations were mutually exclusive.

Conclusions: Mutational screening on fresh cancer cells is an achievable, safe and cost-effective procedure which might allow routinely tumor molecular profiling as powerful integration of conventional histopathological analysis.

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Figures

Fig. 1
Fig. 1
Flow chart of the study design
Fig. 2
Fig. 2
Panel of traces for EGFR and Kras. Panel A: Sequencing chromatograms show (arrows) the deletion in EGFR exon 19 (delE746-L750; K745K) and the EGFR exon 20 T790M plus exon 21 L858R and Kras exon 2 G12V mutations. In the exon 20 chromatogram, the non somatic mutation Q787Q is also present. The percentage of mutant DNA is 10 %. Panel B: Melting peaks of HRM amplicons are compared between wild type (black arrow) and representative mutant samples at different DNA percentage (dashed arrow represents the minimum percentage of 5 %)
Fig. 3
Fig. 3
Comparison between HRM-PCR with “fresh DNA” (A) and “FF-PE DNA” (B)
See this image and copyright information in PMC

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