title: Assessment of EGFR and K-ras mutations in fixed and fresh specimens             from transesophageal ultrasound-guided fine needle aspiration in non-small cell             lung cancer patients.
abstract: In non-small cell lung cancer (NSCLC) patients, somatic EGFR and K-ras mutations predict therapeutic effectiveness and resistance, respectively, to EGFR tyrosine kinase inhibitors (TKIs). Transesophageal ultrasound-guided fine needle aspiration (EUS-FNA) is a validated technique for diagnosis and staging of NSCLC. In the present study, we compared the feasibility and reliability of EGFR and K-ras gene mutation analysis in fixed and fresh mediastinal lymph nodes and extra-lymph nodal samples obtained by EUS-FNA in patients suspicious for NSCLC. Thirty-six patients were enrolled into the study. For each patient, DNA was extracted from both fresh samples and fixed cytological smears. Exons 18-21 of EGFR and exon 2 of K-ras were amplified by PCR and mutation status was determined by direct sequencing and pyrosequencing. All cases were eligible for analysis. NSCLC was diagnosed in 32 patients (25 adenocarcinomas and 7 squamous cell carcinomas) and 4 patients were free of malignancy. Of the 25 patients with adenocarcinoma, EGFR mutations were detected in 2 (8%) fresh tumor samples and in 3 (12%) fixed cytological smears. K-ras mutations were detected in 8 (32%) fresh samples, and in 9 (36%) fixed cytological smears. Fixed and stained cytological samples seem to be more reliable than fresh material for molecular analysis.
authors.affiliation: Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori, Meldola, Italy. p.ulivi@irst.emr.it
authors.forename: Paola
authors.initials: P
authors.lastname: Ulivi
authors.forename: Micaela
authors.initials: M
authors.lastname: Romagnoli
authors.forename: Elisa
authors.initials: E
authors.lastname: Chiadini
authors.forename: Gian-Luca
authors.initials: GL
authors.lastname: Casoni
authors.forename: Laura
authors.initials: L
authors.lastname: Capelli
authors.forename: Carlo
authors.initials: C
authors.lastname: Gurioli
authors.forename: Wainer
authors.initials: W
authors.lastname: Zoli
authors.forename: Luca
authors.initials: L
authors.lastname: Saragoni
authors.forename: Alessandra
authors.initials: A
authors.lastname: Dubini
authors.forename: Anna
authors.initials: A
authors.lastname: Tesei
authors.forename: Dino
authors.initials: D
authors.lastname: Amadori
authors.forename: Venerino
authors.initials: V
authors.lastname: Poletti
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D044963:Biopsy, Fine-Needle; D002289:Carcinoma, Non-Small-Cell Lung / Q000000981:diagnostic imaging / Q000235:genetics* / Q000473:pathology; D004252:DNA Mutational Analysis; D017931:DNA Primers; D019160:Endosonography; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D006801:Humans; D008168:Lung / Q000000981:diagnostic imaging / Q000473:pathology; D008175:Lung Neoplasms / Q000000981:diagnostic imaging / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D011446:Prospective Studies; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins / Q000235:genetics*
chemical_list: D017931:DNA Primers; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D066246:ErbB Receptors; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins