Journal List > Korean J Cytopathol > v.19(2) > 1006516

Korean J Cytopathol. 2008 Sep;19(2):126-135. English.
Published online September 30, 2008.  https://doi.org/10.3338/kjc.2008.19.2.126
Copyright © 2008 The Korean Society for Cytopathology
Methylation Abnormality in Body Fluid Cytology: A Supplemental Molecular Marker for the Diagnosis of Malignant Mesothelioma
Joon Seon Song, M.D., Jin Kyung Jung, M.S., Ji Hye Kang, M.S., Ilseon Hwang, M.D. and Se Jin Jang, M.D., Ph.D.
Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea.
Received July 16, 2008; Accepted August 26, 2008.

Abstract

Malignant mesothelioma (MM) is a highly lethal neoplasm arising in pleura and the peritoneum and a rapid and accurate diagnosis is crucial for treatment of the disease. However, the sensitivity of cytological analysis using pleural or ascitic fluid is relatively low, yielding an accurate diagnosis in only 32~79% of cases. We tested the diagnostic value of epigenetic alterations in body fluid cytology as a supplement to conventional methods. Paraffin-embedded tissue blocks from 21 MM patients and associated body fluid cytology slides considered no evidence of malignancy were used to test for epigenetic alteration. Using methylation-specific PCR, we detected methylation of RASSF1A and p16 in 47.6% (10/21) of both surgically resected tumor samples, respectively. Body fluid samples of MM also showed abnormal methylation of RASSF1A and p16INK4a genes in 38.1% (8/21) and 33.3% (7/21) of cases. The concordance in the rates of RASSF1A and p16INK4a gene-methylation abnormalities determined from cytology samples and tissue samples were 61.9% (13/21) and 66.7% (14/21), respectively. Combining both genes increases the sensitivity of the test to 57.1% (12 of 21) of cases. Our results suggest that testing for methylation abnormalities in selected individual genes or gene combinations has diagnostic value as an alternative or adjunct method to conventional cytological diagnosis.

Keywords: Mesothelioma; Methylation; p16 INK4a; RASSF1A; Body Fluid

Figures


Fig. 1
A. Epithelioid pleural malignant mesothelioma (MM) exhibits tubulopapillary profiles of polygonal cells (patient 8). B. Cytologic specimen of epithelioid pleural MM scored as negative for malignancy and reactive mesothelial hyperplasia. C. An example of sarcomatoid mesothelioma (patient 13) showing atypical spindle cells arranged in fascicles. D. Cytologic specimens from a sarcomatoid pleural MM patient show reactive mesothelial cells. (A,C : H&E, B,D : Papanicolaou stain).
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Fig. 2
Examples of representative MSP analyses of malignant mesothelioma patients, NSCLC controls and inflammatory non-neoplastic controls. Products were amplified with primers that recognize methylated and unmethylated sequences, respectively.
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Tables


Table 1
Clinocopathologic characteristics of patients with malignant mesothelioma and methylation patterns of p16 and RASSF1A genes
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Table 2
Clinocopathologic characteristics of patients with inflammatory/ non-cancer body fluid and Non-small cell lung cancer
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Table 3
PCR primer sequences and PCR product sizes used for MSP
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Notes

This work was supported by KOSEF research grant R01-2004-000-10670-0.

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