Detection of Cell-Free DNA in Bronchial Lavage Fluid Supernatants of Patients with Lung Cancer

doi:10.1196/annals.1318.031

Address for correspondence: M. Fleischhacker, Charité-Universitätsmedizin Berlin, Medizinische Klinik II, Onkologie u. Hämatologie, Mol. Biol. Laboratory, Schumannstr. 20-21, 10117 Berlin. Voice: +49 30 450 51 33 04 michael.fleischhacker{at}charite.de
Ann. N.Y. Acad. Sci. 1022: 202-210 (2004).

Recently, it was shown that it is possible to isolate free circulatingDNA from plasma/serum of patients with benign and malignantdiseases. In addition, several groups were able to detect tumor-associatedalterations in these nucleic acids. We wondered whether anynucleic acids are detectable in cell-free bronchial lavage supernatants,which until now have been discarded after cell harvest. Additionally,we wanted to find out if it is possible to detect tumor-associatedalterations in these DNA molecules. DNA was isolated from cell-freelavage supernatants from 30 lung cancer patients, and the DNAwas examined for microsatellite alterations. Intact DNA couldbe isolated from all cell-free bronchial lavage supernatants.Microsatellite alterations were found in lavage supernatantsof 12 of 30 patients and in lavage cells of 6 of 30 patients.Altogether, alterations were found in 14 of 30 patients. Thus,we could demonstrate for the first time that it is possibleto isolate intact DNA from cell-free bronchial lavage supernatants.Their quantity and quality are sufficient for further amplificationvia polymerase chain reaction. Altogether, tumor-associatedchanges were detected in the DNA of 47% of the patients thatwere analyzed.