title: Determination ofEGFRandKRASmutational status in Greek non-small-cell lung cancer patients.
abstract: It has been reported that certain patients with non-small-cell lung cancer (NSCLC) that harbor activating somatic mutations within the tyrosine kinase domain of theepidermal growth factor receptor(EGFR) gene may be effectively treated using targeted therapy. The use of EGFR inhibitors in patient therapy has been demonstrated to improve response and survival rates; therefore, it was suggested that clinical screening for EGFR mutations should be performed for all patients. Numerous clinicopathological factors have been associated with EGFR andKirsten-rat sarcoma oncogene homolog(KRAS) mutational status including gender, smoking history and histology. In addition, it was reported that EGFR mutation frequency in NSCLC patients was ethnicity-dependent, with an incidence rate of ~30% in Asian populations and ~15% in Caucasian populations. However, limited data has been reported on intra-ethnic differences throughout Europe. The present study aimed to investigate the frequency and spectrum of EGFR mutations in 1,472 Greek NSCLC patients. In addition, KRAS mutation analysis was performed in patients with known smoking history in order to determine the correlation of type and mutation frequency with smoking. High-resolution melting curve (HRM) analysis followed by Sanger sequencing was used to identify mutations in exons 18-21 of the EGFR gene and in exon 2 of the KRAS gene. A sensitive next-generation sequencing (NGS) technology was also employed to classify samples with equivocal results. The use of sensitive mutation detection techniques in a large study population of Greek NSCLC patients in routine diagnostic practice revealed an overall EGFR mutation frequency of 15.83%. This mutation frequency was comparable to that previously reported in other European populations. Of note, there was a 99.8% concordance between the HRM method and Sanger sequencing. NGS was found to be the most sensitive method. In addition, female non-smokers demonstrated a high prevalence of EGFR mutations. Furthermore, KRAS mutation analysis in patients with a known smoking history revealed no difference in mutation frequency according to smoking status; however, a different mutation spectrum was observed.
authors.affiliation: GeneKor, Athens 15344, Greece.
authors.forename: Eirini
authors.initials: E
authors.lastname: Papadopoulou
authors.affiliation: GeneKor, Athens 15344, Greece.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Tsoulos
authors.affiliation: GeneKor, Athens 15344, Greece.
authors.forename: Angeliki
authors.initials: A
authors.lastname: Tsirigoti
authors.affiliation: GeneKor, Athens 15344, Greece.
authors.forename: Angela
authors.initials: A
authors.lastname: Apessos
authors.affiliation: GeneKor, Athens 15344, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Agiannitopoulos
authors.affiliation: GeneKor, Athens 15344, Greece.
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Metaxa-Mariatou
authors.affiliation: Pulmonary Department, Oncology Unit, 'G. Papanikolaou' General Hospital, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Zarogoulidis
authors.affiliation: Pulmonary Department, Oncology Unit, 'G. Papanikolaou' General Hospital, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece.
authors.forename: Pavlos
authors.initials: P
authors.lastname: Zarogoulidis
authors.affiliation: Oncology Unit, General Hospital of Kavala, Kavala 65500, Greece.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Kasarakis
authors.affiliation: Department of Medical Oncology, University General Hospital of Alexandroupolis, Evros 68100, Greece.
authors.forename: Stylianos
authors.initials: S
authors.lastname: Kakolyris
authors.affiliation: Department of Thoracic Surgery, Athens Medical Center, Athens 30606, Greece.
authors.forename: Jubrail
authors.initials: J
authors.lastname: Dahabreh
authors.affiliation: University Clinic of Pulmonology, 'Sotiria' Chest Diseases Hospital, Athens 11527, Greece.
authors.forename: Fotis
authors.initials: F
authors.lastname: Vlastos
authors.affiliation: Department of Oncology, Evaggelismos Hospital, Athens 10676, Greece.
authors.forename: Charalampos
authors.initials: C
authors.lastname: Zoublios
authors.affiliation: Second Pulmonary Clinic, 'Sotiria' Chest Diseases Hospital, Athens 11527, Greece.
authors.forename: Aggeliki
authors.initials: A
authors.lastname: Rapti
authors.affiliation: Fifth Pulmonary Clinic, 'Sotiria' Chest Diseases Hospital, Athens 11527, Greece.
authors.forename: Niki Georgatou
authors.initials: NG
authors.lastname: Papageorgiou
authors.affiliation: Department of Respiratory and Critical Care Medicine (KAA), 'Sotiria' Chest Diseases Hospital, Athens 11527, Greece.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Veldekis
authors.affiliation: Seventh Pulmonary Clinic, 'Sotiria' Chest Diseases Hospital, Athens 11527, Greece.
authors.forename: Mina
authors.initials: M
authors.lastname: Gaga
authors.affiliation: Second Department of Medical Oncology, 'Agii Anargiri' Cancer Hospital, Athens 11524, Greece.
authors.forename: Gerasimos
authors.initials: G
authors.lastname: Aravantinos
authors.affiliation: Department of Medical Oncology, Medical School, Aristotle University, Papageorgiou Hospital, Thessaloniki 54124, Greece.
authors.forename: Vasileios
authors.initials: V
authors.lastname: Karavasilis
authors.affiliation: Second Department of Respiratory Medicine, Sismanoglio-A. Fleming General Hospital of Attiki, Athens 15126, Greece.
authors.forename: Napoleon
authors.initials: N
authors.lastname: Karagiannidis
authors.affiliation: GeneKor, Athens 15344, Greece.
authors.forename: George
authors.initials: G
authors.lastname: Nasioulas
keywords: Kirsten-rat sarcoma oncogene homolog; Sanger sequencing; epidermal growth factor receptor; high-resolution melting curve analysis; next generation sequencing; non-small-cell lung cancer