Detection of cell-free DNA in bronchial lavage fluid supernatants of patients with lung cancer.
Recently, it was shown that it is possible to isolate free circulating DNA from plasma/serum of patients with benign and malignant diseases. In addition, several groups were able to detect tumor-associated alterations in these nucleic acids. We wondered whether any nucleic 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-associated alterations in these DNA molecules. DNA was isolated from cell-free lavage supernatants from 30 lung cancer patients, and the DNA was examined for microsatellite alterations. Intact DNA could be isolated from all cell-free bronchial lavage supernatants. Microsatellite alterations were found in lavage supernatants of 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 possible to isolate intact DNA from cell-free bronchial lavage supernatants. Their quantity and quality are sufficient for further amplification via polymerase chain reaction. Altogether, tumor-associated changes were detected in the DNA of 47% of the patients that were analyzed.
Carstensen
Tim
T
Charité-Universitätsmedizin Berlin, Medizinische Klinik II, Pulmologie, Berlin, Germany.
Schmidt
Bernd
B
Engel
Eileen
E
Jandrig
Burkhard
B
Witt
Christian
C
Fleischhacker
Michael
M
D000368:Aged; D014408:Biomarkers, Tumor; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis / Q000235:genetics; D018288:Carcinoma, Small Cell / Q000175:diagnosis / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics* / Q000302:isolation & purification; D005240:Feasibility Studies; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D019656:Loss of Heterozygosity; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D018895:Microsatellite Repeats; D008875:Middle Aged; D016133:Polymerase Chain Reaction
D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm