This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ferretti, G. Articles by Goldhirsch, A. Search for Related Content PubMed PubMed Citation Articles by Ferretti, G. Articles by Goldhirsch, A.

Detection by Denaturant Gradient Gel Electrophoresis of Tumor-specific Mutations in Biopsies and Relative Bronchoalveolar Lavage Fluid from Resectable Non-Small Cell Lung Cancer

Divisions of Medical Oncology [G. Fe., G. C., T. D. P., L. O., M. M., M. C., F. d. B., N. F., A. G.] and Thoracic Surgery [U. P., L. S.], European Institute of Oncology, 20141 Milano, Italy; and Institute of General Pathology [A. C., G. Fl.], Institute of Anesthesiology and Intensive Care Medicine [M. G. C.], Division of Thoracic Surgery [P. L. G.], and Division of Internal Medicine [G. P.], Catholic University, School of Medicine, 00168 Rome, Italy

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.

This article has been cited by other articles:

				J Peltonen, J A Welsh, and K H Vahakangas Is there a role for PCR-SSCP among the methods for missense mutation detection of TP53 gene? Human and Experimental Toxicology, January 1, 2007; 26(1): 9 - 18. [Abstract] [PDF]

				M. S. Brose, P. Volpe, M. Feldman, M. Kumar, I. Rishi, R. Gerrero, E. Einhorn, M. Herlyn, J. Minna, A. Nicholson, et al. BRAF and RAS Mutations in Human Lung Cancer and Melanoma Cancer Res., December 1, 2002; 62(23): 6997 - 7000. [Abstract] [Full Text] [PDF]

				P. S. Bernard and C. T. Wittwer Real-Time PCR Technology for Cancer Diagnostics Clin. Chem., August 1, 2002; 48(8): 1178 - 1185. [Abstract] [Full Text] [PDF]

				H. Millward, W. Samowitz, C. T. Wittwer, and P. S. Bernard Homogeneous Amplification and Mutation Scanning of the p53 Gene Using Fluorescent Melting Curves Clin. Chem., August 1, 2002; 48(8): 1321 - 1328. [Abstract] [Full Text] [PDF]