Microsatellite analysis of induced sputum DNA in patients with lung cancer in heavy smokers and in healthy subjects

Abnormality in the fragile histidine triade (FHIT), a candidate tumor suppressor gene located in chromosome region 3 (3p14.2), has been frequently found in multiple tumor types, including lung cancer. In this study, the authors assessed the consistency of DNA microsatellite analysis of induced sputum (IS), as compared to that of blood and plasma. They also evaluated the loss of heterozigosity (LOH) and microsatellite instability (MI) in 3 different loci, D3S1300, D3S1313, and D3S1234, all internal to the FHIT gene, in IS, blood, and plasma from patients with lung cancer, smokers, and healthy subjects. Eighteen patients with lung cancer (3 females, age mean +/- SD: 63 +/- 7years), 39 smokers (23 females, age mean +/- SD: 57 +/- 6 years and cigarette pack-years mean +/- SD: 34 +/- 12), and 22 health nonsmoking subjects (13 females, age mean +/- SD: 63 +/- 5 Years) were studied. DNA was extracted from blood, plasma, and IS, by means of a standard method. Analysis of LOH and MSI were performed using a fluorescent polymerase chain reaction (PCR)-based approach, followed by capillary electrophoresis. The ratios between the peak heights (PHS), expressed as random fluorescence units, from plasma/blood (p/b) and induced sputum/blood. (is/b) in all three loci were considered. The biases (agreement limits) between the mean PH ratio from p/b and is/b of D3S1300, D3S1313, and D3S1234 were respectively 0.07 (-0.39 to 0.53), 0.016 (-0.32 to 0.35), - 0.10 (-0.51 to 0.30) in the patients; - 0.04 (-0.52 to 0.43), - 0.06 (-0.31 to 0.18), - 0.08 (-0.48 to 0.30) in smokers; and - 0.11 (-0.40 to 0.17), -0.05 (-0.53 to 0.43), -0.09 (-0.51 to 0.33) in healthy subjects. LOH and MSI in at least one locus were observed in 55% of patients, in 18% of smokers, and in 4.5% of healthy subjects (P < 0.001). These results showed that IS DNA provided data that were consistent with those from blood and plasma. These findings highlight new prospects for early tumor detection by a noninvasive technique based on the analysis of genetic alterations in induced sputum.