Detection by denaturant gradient gel electrophoresis of tumor-specific mutations in biopsies and relative bronchoalveolar lavage fluid from resectable non-small cell lung cancer.
A PCR-denaturant gradient gel electrophoresis (DGGE) method was developed for the detection of p53 and K-ras mutations in primary operable tumors and paired BAL samples of non-small cell lung cancer. Among 36 patients, 9 showed p53 exon V mutations in biopsies and in three paired bronchoalveolar lavage (BAL) specimens with a 33% concordance. Five patients presented p53 exon VI mutations in biopsies and in two paired BALs with a 40% concordance. No mutations were found in p53 exon VII either in biopsies or in paired BAL samples with 100% concordance. Exon VIII mutations were found in six primary tumors and in two BALs with a 33% concordance. Of 36 patients, we detected 7 (19.4%) with K-ras exon I mutations on tumor samples. DGGE analysis of DNA from BAL samples revealed three mutations distributed on K-ras exon I with a 42% overall concordance with respect to tumor tissue. Molecular screening by DGGE of p53-amplified DNA from BAL had cumulative 46.6% sensitivity, 100% specificity, and 77.7% accuracy. DGGE K-ras detection showed 43% sensitivity, 100% specificity, and 88.8% test accuracy. The method proposed demonstrated to be specific, accurate, and at relatively low cost but limited by low sensitivity in detecting the presence of neoplastic cells in patients with resectable non-small cell lung cancer.
Ferretti
G
G
Division of Medical Oncology, European Institute of Oncology, Milano, Italy. gianluigi.ferreti@ieo.it
Curigliano
G
G
Pastorino
U
U
Cittadini
A
A
Flamini
G
G
Calabrò
M G
MG
De Pas
T
T
Orlando
L
L
Mandalà
M
M
Colleoni
M
M
Spaggiari
L
L
Granone
P L
PL
Pagliari
G
G
de Braud
F
F
Fazio
N
N
Goldhirsch
A
A
D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism; D000368:Aged; D001706:Biopsy; D001992:Bronchoalveolar Lavage Fluid; D018196:Carcinoma, Adenosquamous / Q000235:genetics / Q000378:metabolism; D018287:Carcinoma, Large Cell / Q000235:genetics / Q000378:metabolism; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000378:metabolism* / Q000601:surgery; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods; D004586:Electrophoresis / Q000379:methods*; D005091:Exons; D005260:Female; D016158:Genes, p53 / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000601:surgery; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016283:Proto-Oncogene Proteins p21(ras) / Q000096:biosynthesis / Q000235:genetics; D012680:Sensitivity and Specificity; D012907:Smoking; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis
D016159:Tumor Suppressor Protein p53; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)