Tuberc Respir Dis.  2008 Apr;64(4):285-292. 10.4046/trd.2008.64.4.285.

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

• Rha SH
• Lee SM
• Koo TH
• Shin BC
• Huh JH
• Um SJ
• Yang DK
• Lee SK
• Son C
• Roh MS
• Bae HJ
• Kim KN
• Lee KN
• Choi PJ

• KMID: 1970179
• DOI: http://doi.org/10.4046/trd.2008.64.4.285

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.

Keyword

Pleural effusion; Malignancy; Hypermethylation

MeSH Terms

Biopsy
Genes, p16
Humans
Lung Neoplasms
Methylation
Pleural Effusion
Pleural Effusion, Malignant
Polymerase Chain Reaction
Prognosis
Receptors, Retinoic Acid
Smoke
Receptors, Retinoic Acid
Smoke

Figure

• 

  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.

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