title: Assessment of epidermal growth factor receptor and K-ras mutation status in cytological stained smears of non-small cell lung cancer patients: correlation with clinical outcomes.
abstract: OBJECTIVE
  Epidermal growth factor receptor (EGFR) and K-ras mutations guide treatment selection in non-small cell lung cancer (NSCLC) patients. Although mutation status is routinely assessed in biopsies, cytological specimens are frequently the only samples available. We determined EGFR and K-ras mutations in cytological samples.
  
  
  METHODS
  DNA was extracted from 150 consecutive samples, including 120 Papanicolau smears (80%), 10 cell blocks (7%), nine fresh samples (6%), six ThinPrep® tests (4%), and five body cavity fluids (3.3%). Papanicolau smears were analyzed when they had >50% malignant cells. Polymerase chain reaction and direct sequencing of exons 18-21 of EGFR and exon 2 of K-ras were performed. EGFR mutations were simultaneously determined in biopsies and cytological samples from 20 patients. Activity of EGFR tyrosine kinase inhibitors (TKIs) was assessed.
  
  
  RESULTS
  The cytological diagnosis was adenocarcinoma in 110 samples (73%) and nonadenocarcinoma in 40 (27%) samples. EGFR mutations were identified in 26 samples (17%) and K-ras mutations were identified in 18 (12%) samples. EGFR and K-ras mutations were mutually exclusive. In EGFR-mutated cases, DNA was obtained from stained smears in 24 cases (92%), pleural fluid in one case (4%), and cell block in one case (4%). The response rate to EGFR TKIs in patients harboring mutations was 75%. The mutation status was identical in patients who had both biopsies and cytological samples analyzed.
  
  
  CONCLUSIONS
  Assessment of EGFR and K-ras mutations in cytological samples is feasible and comparable with biopsy results, making individualized treatment selection possible for NSCLC patients from whom tumor biopsies are not available.
authors.affiliation: Department of Pathology, University Clinic of Navarra, University of Navarra, Pamplona, Spain. mdlozano@unav.es
authors.forename: Maria D
authors.initials: MD
authors.lastname: Lozano
authors.forename: Javier J
authors.initials: JJ
authors.lastname: Zulueta
authors.forename: Jose I
authors.initials: JI
authors.lastname: Echeveste
authors.forename: Alfonso
authors.initials: A
authors.lastname: Gúrpide
authors.forename: Luis M
authors.initials: LM
authors.lastname: Seijo
authors.forename: Salvador
authors.initials: S
authors.lastname: Martín-Algarra
authors.forename: Anabel
authors.initials: A
authors.lastname: Del Barrio
authors.forename: Ruben
authors.initials: R
authors.lastname: Pio
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: Idoate
authors.forename: Tania
authors.initials: T
authors.lastname: Labiano
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Perez-Gracia
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy* / Q000235:genetics; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000970:Antineoplastic Agents / Q000627:therapeutic use; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000235:genetics; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000302:isolation & purification; D004791:Enzyme Inhibitors / Q000627:therapeutic use; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000235:genetics*; D005091:Exons; D005260:Female; D011905:Genes, ras; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016133:Polymerase Chain Reaction; D013194:Staining and Labeling; D016896:Treatment Outcome; D018631:ras Proteins / Q000235:genetics*
chemical_list: D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D004791:Enzyme Inhibitors; D066246:ErbB Receptors; D018631:ras Proteins