p16INK4a Promoter Hypermethylation in Sputum, Blood, and Tissue from Non-Small Cell Lung Cancer and Pulmonary Inflammation

Jeong Pyo Kim; Kyong Mee Kim; Soon Seog Kwon; Young Kyoon Kim; Kwan Hyoung Kim; Hwa Sik Moon; Jeong Sup Song; Sung Hak Park; Joong Hyun Ahn

doi:10.4046/trd.2006.60.02.160

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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

Original Article

Tuberc Respir Dis 2006;60(2):160-170.

Published online: 19 February 2006

DOI: https://doi.org/10.4046/trd.2006.60.02.160

p16^INK4a Promoter Hypermethylation in Sputum, Blood, and Tissue from Non-Small Cell Lung Cancer and Pulmonary Inflammation

Jeong Pyo Kim, M.D., Kyong Mee Kim, M.D.¹, Soon Seog Kwon, M.D., Young Kyoon Kim, M.D., Kwan Hyoung Kim, M.D., Hwa Sik Moon, M.D., Jeong Sup Song, M.D., Sung Hak Park, M.D., Joong Hyun Ahn, M.D.

Department of Internal Medicine, Department of Pathology

¹College of Medicine, the Catholic University of Korea, Seoul, Korea

Address for correspondence : Joong Hyun Ahn, M.D. Department of Internal Medicine, Our Lady of Mercy Hospital, College of Medicine, the Catholic University of Korea, 665, Bupyong-dong, Bupyong-gu, Incheon, 403-720, Korea Phone : 032-510-5500 Fax : 032-510-5683 E-mail : ajhcmcim@olmh.cuk.ac.kr

Received 4 October 2005 Accepted 19 June 2006

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 p16^INK4a 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 p16^INK4amethylation was determined using methylation-specific polymerase chain reaction.

Results :

p16^INK4a 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 p16^INK4a 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 p16^INK4a promoter hypermethylation of the blood can used to discriminate between NSCLC and pulmonary inflammation.

Keywords: p16^INK4a promoter hypermethylation, Non-small cell lung cancer, 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.

Figure 2.

Analysis of p16INK4a promoter hypermethylation in sputum from NSCLC and pulmonary inflammation.

Figure 3.

Analysis of p16INK4a promoter hypermethylation in tissue from NSCLC. Methylation was observed in six of eight cases.

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.

p16^INK4a 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 p16^INK4a 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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