Journal List > Tuberc Respir Dis > v.64(4) > 1001218

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Rha, Lee, Koo, Shin, Huh, Um, Yang, Lee, Son, Roh, Bae, Kim, Lee, and Choi: Diagnosis of Malignant Pleural Effusion by using Aberrant Methylation of p16 and RARB2
Original Article
Tuberculosis and Respiratory Diseases

Tuberculosis and Respiratory Diseases 2008; 64(4): 285-292.

Published online: 30 April 2008

DOI: https://doi.org/10.4046/trd.2008.64.4.285

Diagnosis of Malignant Pleural Effusion by using Aberrant Methylation of p16 and RARB2

Seo Hee Rha, M.D.2, Su Mi Lee, M.D.1, Tae Hyoung Koo, M.D.1, Bong Chul Shin, M.D.1, Jung Hun Huh, M.D.1, Soo Jung Um, M.D.1, Doo Kyung Yang, M.D.1, Soo-Keol Lee, M.D.1, Choonhee Son, M.D.1, Mee Sook Roh, M.D.2, Ho-Jeong Bae2, Ki Nam Kim, M.D.3, Ki Nam Lee, M.D.3, Pil Jo Choi, M.D.4

1Department of Internal Medicine, Dong-A University College of Medicine, Busan, Korea.

2Department of Pathology, Dong-A University College of Medicine, Busan, Korea.

3Department of Radiology, Dong-A University College of Medicine, Busan, Korea.

4Department of Thoracic and Cardiovascular Surgery, Dong-A University College of Medicine, Busan, Korea.

Address for correspondence: Choonhee Son, M.D. Department of Internal Medicine, Dong-A University College of Medicine, 1, Dongdaesin-dong, Seo-gu, Busan 602-714, Korea. Phone: 82-51-240-2874, Fax: 82-51-242-5852, son30243@hanmail.net

Received 14 February 2008       Accepted 28 March 2008

Copyright © 2008 The Korean Academy of Tuberculosis and Respiratory Diseases

Abstract

Background

A diagnosis of malignant pleural effusion is clinically important, as the prognosis of lung cancer patients with malignant pleural effusion is poor. The diagnosis will be difficult if a cytological test is negative. This study was performed to investigate whether the detection of hypermethylation of the p16 (CDKN2A) and retinoic acid receptor b2 (RARB2) genes in pleural fluid is useful for a diagnosis of malignant pleural effusion.

Methods

Pleural effusion was collected from 43 patients and was investigated for the aberrant promoter methylation of the RARB2 and CDKN2A genes by use of methylation-specific PCR. Results were compared with findings from a pleural biopsy and from pleural fluid cytology.

Results

Of 43 cases, 17 cases of pleural effusion were due to benign diseases, and 26 cases were from lung cancer patients with malignant pleural effusion. Hypermethylation of the RARB2 and CDKN2A genes was not detected in the case of benign diseases, independent of whether or not the patients had ever smoked. In 26 cases of malignant pleural effusion, hypermethylation of RARB2, CDKN2A or either of these genes was detected in 14, 5 and 15 cases, respectively. The sensitivities of a pleural biopsy, pleural fluid cytology, hypermethylation of RARB2, hypermethylation of CDKN2A, or hypermethylation of either of the genes were 73.1%, 53.8%, 53.8%, 19.2%, and 57.7%, respectively; negative predictive values were 70.8%, 58.6%, 58.6%, 44.7%, and 60.7%, respectively. If both genes are considered together, the sensitivity and negative predictive value was lower than that for a pleural biopsy, but higher than that for pleural fluid cytology. The sensitivity of hypermethylation of the RARB2 gene for malignant pleural effusion was lower in small cell lung cancers than in non-small cell lung cancers.

Conclusion

These results demonstrate that detection of hypermethylation of the RARB2 and CDKN2A genes showed a high specificity, and sensitivity was higher than for pleural fluid cytology. With a better understanding of the pathogenesis of lung cancer according to histological types at the molecular level, and if appropriate genes are selected for hypermethylation testing, more precise results may be obtained.

Keywords: Pleural effusion, Malignancy, Hypermethylation

Figures and Tables

Figure 1
The results of methylation specific PCR of retinoic acid receptor beta (A) and p16 (B) in cells collected from the pleural effusion of lung cancer patients (24~35) and control (1~5). Both unmethylated (U) and methylated (M) PCR products are shown for each sample. *patient number DNA ladder, †positive control, ‡normal saline for negative control, §methylaed, ∥unmethylated.
trd-64-285-g001
Table 1
Primers used in methylation specific PCR
trd-64-285-i001

*product size, †annealing temperature, ‡retinoic acid receptor beta 2.

Table 2
The results of the aberrant methylation and the characteristics of the patients
trd-64-285-i002

*female, †male, ‡non-small cell lung cancer, §negative for aberrant methylation, ∥positive for aberrant methylation, retinoic acid receptor beta 2, **negative for malignancy in histology or cytology, ††positive for malignancy in histology or cytology.

Table 3
The sensitivity and negative predictive values for the detection of malignant pleural effusion according to the detecting methods
trd-64-285-i003

*retinoic acid receptor beta 2, †adenocarcinoma, ‡squamous cell carcinoma, §small cell carcinoma, ∥unclassifiable non-small cell lung cancer, negative predictive value.

Table 4
The sensitivity for the detection of malignant pleural effusion by aberrant methylation according to the smoking status
trd-64-285-i004

*retinoic acid receptor beta 2.

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