Journal List > Tuberc Respir Dis > v.60(2) > 1000905

Kim, Kim, Kwon, Kim, Kim, Moon, Song, Park, and Ahn: p16INK4a Promoter Hypermethylation in Sputum, Blood, and Tissue from Non-Small Cell Lung Cancer and Pulmonary Inflammation

Abstract

Background :

The aberrant promoter hypermethylation of p16INK4a, as a tumor suppressor gene, is contributory factor to non-small cell lung cancer(NSCLC). However, its potential diagnostic impact of lung cancer is unclear. This study measured the level of p16INK4a promoter hypermethylation in the sputum and blood, and compared this with the level measured in the tissue obtained from NSCLC and pulmonary inflammation.

Methods :

Of the patients who visited the Our Lady of Mercy Hospital in Incheon, Korea for an evaluation of a lung mass and underwent blood, sputum, and tissue tests, 23patients (18 NSCLC, 5 pulmonary inflammation) were enrolled in this study. DNA was extracted from each sample and the level of p16INK4amethylation was determined using methylation-specific polymerase chain reaction.

Results :

p16INK4a methylation of the blood was observed in 88.9% (16 of 18) and 20.0% (1 of 5) of NSCLC and from pulmonary inflammation samples, respectively (P=0.008). Methylation of the sputum was observed in 83.3% (10 of 12) 80.0% (4 of 5) of NSCLC and pulmonary inflammation samples, respectively (P=1.00). Among the 8 NSCLC tissue samples, methylation changes were detected in 75.0% of samples (6 cases). Four out of seven tissue samples (57.1%) showed concordance, being methylated in both the blood and sputum.

Conclusions :

There was a higher level of p16INK4a methylation of the blood from NSCLC patients than from pulmonary inflammation. The tissue showed a high concordance with the blood in the NSCLC samples. These findings suggest that p16INK4a promoter hypermethylation of the blood can used to discriminate between NSCLC and pulmonary inflammation.

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Figure 1.
Analysis of p16INK4a promoter hypermethylation in blood from NSCLC and pulmonary inflammation. Bisulfite modified DNA was amplified by primers specific to unmethylated or methylated alleles of p16INK4a. The prescence of visible PCR product in each lanes marked unmethylated or methylated.
trd-60-160f1.tif
Figure 2.
Analysis of p16INK4a promoter hypermethylation in sputum from NSCLC and pulmonary inflammation.
trd-60-160f2.tif
Figure 3.
Analysis of p16INK4a promoter hypermethylation in tissue from NSCLC. Methylation was observed in six of eight cases.
trd-60-160f3.tif
Table 1.
Clinical characteristics of patients
Characteristic NSCLC Pulmonary inflammation P value
No. of cases 18 5  
Age (yr) 68±8 65±10 0.549
Sex (no. of cases)      
Male 16 4 0.539
Female 2 1 0.539
Smoking history      
Pack-years 39±25 24±25 0.329
Current (no. of cases) 12 1  
Former 4 2  
Years since quitting 10±13 10±13  
Never 2 2 4

Plus-minus values are mean±SD.

NSCLC: non-small cell lung cancer

Table 2.
p16INK4a hypermethylation in NSCLC and pulmonary inflammation
Sample (no. of cases) NSCLC Pulmonary inflmmation P value
Blood 16/18 (88.9%) 1/5 (20.0%) 0.008
Sputum 10/12 (83.3%) 4/5 (80.0%) 1.00
Tissue 6/8 (75.0%)

NSCLC: non‐small cell lung cancer

Table 3.
Combined p16INK4a hypermethylation in blood, sputum, and tissue from NSCLC
Methylation status No. of cases %
T+    
B+S+ 4 57.1
B+S‐ 1 14.3
T‐    
B+S+ 1 14.3
B+S‐ 1 14.3
Total 7 100

NSCLC: non‐small cell lung cancer

T: tissue, B: blood, S: sputum

‡One case of total 8 tissue was excluded, since the one's sputum DNA could not be acquired.

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