title: Automated real-time PCR to determine K-ras codon 12 mutations in non-small cell lung cancer: comparison with immunohistochemistry and clinico-pathological features.
abstract: A real-time PCR technique with automated computerized analysis (TaqMan ) was tested to detect K-ras mutations in 66 patients suffering from NSCLC. This technology is characterized by high reproducibility of data and a time-saving analysis procedure. In 11% (7/66) of the tumour specimens and 2% (1/58) of adjacent tumour-free lung specimens a K-ras codon 12 mutation was detected. In adenocarcinomas containing > or =40% tumour cells, however, K-ras mutations were seen in 25% of the cases. The point mutations detected in tumours were GGT right curved arrow TGT in five cases and GGT right curved arrow GTT in two cases. As compared with immunohistochemical parameters, the K-ras mutated group was characterized by a c-erbB-2 negativity (p=0.04) and a smaller number of c-erbB-3 (p=0.02) positive cases. EGFR, bcl-2, p53, Ki-67 and p120 expression did not differ significantly. Determination of the K-ras point mutations by automated TaqMan PCR in NSCLC tumour specimens is feasable and highly specific. Due to its high throughput capacity this method represents a valuable tool for routine screening.
authors.affiliation: Department of Internal Medicine, Innsbruck University Hospital, A-6020 Innsbruck, Austria. wolfgang.hilbe@uibk.ac.at
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hilbe
authors.forename: Margit
authors.initials: M
authors.lastname: Dlaska
authors.forename: Hans-Christoph
authors.initials: HC
authors.lastname: Duba
authors.forename: Stephan
authors.initials: S
authors.lastname: Dirnhofer
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Eisterer
authors.forename: Frieda
authors.initials: F
authors.lastname: Oberwasserlechner
authors.forename: Andreas
authors.initials: A
authors.lastname: Mildner
authors.forename: Thomas
authors.initials: T
authors.lastname: Schmid
authors.forename: Thomas
authors.initials: T
authors.lastname: Kühr
authors.forename: Ewald
authors.initials: E
authors.lastname: Wöll
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D045744:Cell Line, Tumor; D003062:Codon; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D008168:Lung / Q000473:pathology; D008175:Lung Neoplasms / Q000235:genetics*; D009154:Mutation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D003062:Codon