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FISH Assay Development For The Detection Of P16/CDKN2A Deletion In Malignant Pleural Mesothelioma
C. Chung, G. Santos, D. Hwang, O. Ludkovski, M. Pintilie, J. Squire, M. Tsao
Published 2010 · Medicine, Biology
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Aims To develop a fluorescence in-situ hybridisation (FISH) assay for detecting p16/CDKN2A deletion on paraffin tissue sections for use as an ancillary test to distinguish reactive from malignant mesothelial proliferations. Method Dual-colour FISH for p16/CDKN2A and chromosome 9 (CEP-9) was performed on 11 benign mesothelial proliferations and 54 malignant pleural mesothelioma (MPM) cases to establish cut-off values for p16/CDKN2A deletion. A third MYC probe was used to verify cases showing homozygous deletion. Eight equivocal biopsies were used for assay testing. Results Cut-off values for p16/CDKN2A deletion were calculated based on FISH signalling patterns obtained from the benign controls (mean percent nuclei plus three standard deviations). Hemizygous deletion was defined as >44% of nuclei showing the hemizygous (one p16/CDKN2A, two CEP-9 signals) or >15% of nuclei showing the monosomy (one p16/CDKN2A, one CEP-9 signal) deletion patterns. None of the benign cases showed a homozygous deletion pattern (no p16/CDKN2A, at least one CEP-9 signal). In the malignant cases, the percentage of nuclei showing homozygous deletion ranged from 1% to 87%. Therefore, the cut-off value for homozygous deletion was defined as >10%. P16/CDKN2A deletion was detected in 61% (33/54) of MPM cases. Among the equivocal biopsies, four showed homozygous and one showed hemizygous p16/CDKN2A deletion. Age over 60 years, asbestos exposure and p16/CDKN2A deletion were associated with a worse prognosis. Conclusion Distinction between benign and malignant mesothelial proliferations can be diagnostically challenging. FISH for p16/CDKN2A deletion is a useful test for confirming the diagnosis of MPM.
This paper references
The use of epithelial membrane antigen and silver‐stained nucleolar organizer regions testing in the differential diagnosis of mesothelioma from benign reactive mesothelioses
K. Metze (1999)
A cell cycle regulator potentially involved in genesis of many tumour types
A. Kamb (1994)
The use of immunohistochemistry in distinguishing reactive from neoplastic mesothelium. A novel use for desmin and comparative evaluation with epithelial membrane antigen, p53, platelet‐derived growth factor‐receptor, P‐glycoprotein and Bcl‐2
R. Attanoos (2003)
Diagnostic importance of 9p21 homozygous deletion in malignant mesotheliomas
S. Chiosea (2008)
The gene for the cyclin‐dependent‐kinase‐4 inhibitor, CDKN2A, is preferentially deleted in malignant mesothelioma
J. Prins (1998)
The use of histological and immunohistochemical markers to distinguish pleural malignant mesothelioma and in situ mesothelioma from reactive mesothelial hyperplasia and reactive pleural fibrosis
P. Cury (1999)
p 16 alterations and deletion mapping of 9 p 21p 22 in malignant mesothelioma
JQ Cheng (1994)
Immunohistochemical detection of GLUT-1 can discriminate between reactive mesothelium and malignant mesothelioma
Y. Kato (2007)
Specificity of interphase fluorescence in situ hybridization for detection of chromosome aberrations in tumor pathology.
M. Tibiletti (2004)
The use of CDKN2A deletion as a diagnostic marker for malignant mesothelioma in body cavity effusions
P. Illei (2003)
Homozygous deletion of CDKN2A and codeletion of the methylthioadenosine phosphorylase gene in the majority of pleural mesotheliomas.
P. Illei (2003)
Immunoreactivity for the β chain of the platelet‐derived growth factor receptor in malignant mesothelioma and non‐neoplastic mesothelium
M. Ramael (1992)
FISH analysis for the detection of lymphoma-associated chromosomal abnormalities in routine paraffin-embedded tissue.
R. A. Ventura (2006)
Alterations of the p16(INK4) locus in human malignant mesothelial tumors.
T. Hirao (2002)
Codeletion of p15 and p16 in primary malignant mesothelioma.
S. Xio (1995)
Wnt pathway activation in mesothelioma: evidence of Dishevelled overexpression and transcriptional activity of beta-catenin.
K. Uematsu (2003)
Sensitivity and specificity of immunohistochemical antibodies used to distinguish between benign and malignant pleural disease: a systematic review of published reports. Histopathology 2006;49:561e8
J King (2006)
The Separation of Benign and Malignant Mesothelial Proliferations
A. Churg (2012)
p16 alterations and deletion mapping of 9p21-p22 in malignant mesothelioma.
J. Cheng (1994)
Cadherins, catenins and APC in pleural malignant mesothelioma
A. S. Abutaily (2003)
p53 immunostaining in the differentiation of reactive processes from malignancy in pleural biopsy specimens.
P. Cagle (1994)
The expression pattern of β‐catenin in mesothelial proliferative lesions and its diagnostic utilities
Yiran Dai (2005)
p53 immunostaining in the distinction between benign and malignant mesothelial proliferations using formalin‐fixed paraffin sections
F. Mayall (1992)
Global gene expression profiling of pleural mesotheliomas: overexpression of aurora kinases and P16/CDKN2A deletion as prognostic factors and critical evaluation of microarray-based prognostic prediction.
Fernando López-Ríos (2006)
Sensitivity and specificity of immunohistochemical antibodies used to distinguish between benign and malignant pleural disease: a systematic review of published reports
J. King (2006)
A cell cycle regulator potentially involved in genesis of many tumor types.
A. Kamb (1994)
Immunohistochemical analysis still has a limited role in the diagnosis of malignant mesothelioma. A study of thirteen antibodies.
F. Roberts (2001)
P16 Loss and Mitotic Activity Predict Poor Survival in Patients with Peritoneal Malignant Mesothelioma
A. Borczuk (2005)
Prognostic significance of p16/cdkn2a loss in pleural malignant mesotheliomas
S. Dacic (2008)
Immunoreactivity for p‐170 glycoprotein in malignant mesothelioma and in non‐neoplastic mesothelium of the pleura using the murine monoclonal antibody JSB‐1
M. Ramael (1992)
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The pathological and molecular diagnosis of malignant pleural mesothelioma: a literature review.
G. Alí (2018)
Prognostic factors in malignant pleural mesothelioma.
B. Davidson (2015)
Mesothelioma patient derived tumor xenografts with defined BAP1 mutations that mimic the molecular characteristics of human malignant mesothelioma
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Tumor Microenvironment: Recent Advances
Wim E Crusio (2020)
The importance of FISH signal cut-off values for 9p21 deletion in malignant pleural mesothelioma: Is it underestimated?
Gamze Kulduk (2019)
Current applications of molecular testing on body cavity fluids.
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Utility of methylthioadenosine phosphorylase immunohistochemical deficiency as a surrogate for CDKN2A homozygous deletion in the assessment of adult-type infiltrating astrocytoma
K. Satomi (2020)
Cryobiopsy during flex-rigid pleuroscopy: an emerging alternative biopsy method in malignant pleural mesothelioma. A comparative study of pathology.
T. Nakai (2019)
Association Between Clinicopathological Factors and Genomic Abnormalities Detected by FISH Analysis in Epithelioid Diffuse Malignant Pleural Mesothelioma
M. Takeda (2017)
Coexistence of p16/CDKN2A homozygous deletions and activating EGFR mutations in lung adenocarcinoma patients signifies a poor response to EGFR-TKIs.
Juhong Jiang (2016)
Morphological difference between pleural mesothelioma cells in effusion smears with either BAP1 loss or 9p21 homozygous deletion and reactive mesothelial cells without the gene alterations
S. Matsumoto (2019)
Usefulness of p16/CDKN2A fluorescence in situ hybridization and BAP1 immunohistochemistry for the diagnosis of biphasic mesothelioma.
D. Wu (2017)
Use of multicolor fluorescence in situ hybridization to detect deletions in clinical tissue sections
Maisa Yoshimoto (2018)
Challenges and controversies in the diagnosis of mesothelioma: Part 1. Cytology-only diagnosis, biopsies, immunohistochemistry, discrimination between mesothelioma and reactive mesothelial hyperplasia, and biomarkers
D. Henderson (2013)
Switching off malignant mesothelioma: exploiting the hypoxic microenvironment
Noushin Nabavi (2016)
Pleural Malignant Mesothelioma : Signi fi cant Advances Histologic Subtyping of Epithelioid Mesothelioma
F. Galateau-Sallé (2016)
Genomic Deletion of BAP1 and CDKN2A Are Useful Markers for Quality Control of Malignant Pleural Mesothelioma (MPM) Primary Cultures
K. Sarun (2018)
CDKN2A copy number and p16 expression in malignant pleural mesothelioma in relation to asbestos exposure
E. Kettunen (2019)
Morphological Characteristics of 9p21 Homozygous Deletion-Positive Malignant Pleural Mesothelioma Cells Detected by FISH
K. Nabeshima (2012)
Iron addiction with ferroptosis‐resistance in asbestos‐induced mesothelial carcinogenesis: Toward the era of mesothelioma prevention
S. Toyokuni (2019)
Patient-Derived Xenograft Establishment from Human Malignant Pleural Mesothelioma
Licun Wu (2016)
Hemizygous loss of NF2 detected by fluorescence in situ hybridization is useful for the diagnosis of malignant pleural mesothelioma
Y. Kinoshita (2019)
Diagnostic utility of BAP1 and EZH2 expression in malignant mesothelioma
A. Shinozaki-Ushiku (2017)
A Rare Case of Mesothelioma Showing Micropapillary and Small Cell Differentiation with Aggressive Behavior
Y. Cha (2014)
Deletion status of p16 in effusion smear preparation correlates with that of underlying malignant pleural mesothelioma tissue
T. Hida (2015)
Genomic Characterization of Pleural Solitary Fibrous Tumours
Ghassan Allo (2013)
A combination of MTAP and BAP1 immunohistochemistry is effective for distinguishing sarcomatoid mesothelioma from fibrous pleuritis.
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