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Jonathan I Epstein - One of the best experts on this subject based on the ideXlab platform.

Tomoyuki Hida - One of the best experts on this subject based on the ideXlab platform.

  • highly expressed ezh2 in combination with bap1 and mtap loss as detected by immunohistochemistry is useful for differentiating malignant pleural mesothelioma from reactive Mesothelial Hyperplasia
    Lung Cancer, 2019
    Co-Authors: Masayo Yoshimura, Tomoyuki Hida, Shinji Matsumoto, Makoto Hamasaki, Akinori Iwasaki, Yoshiaki Kinoshita, Yoshinao Oda, Kazuki Nabeshima
    Abstract:

    Abstract Objective Malignant pleural mesothelioma (MPM) is an aggressive neoplasm with poor prognosis. Loss of BRCA-associated protein 1 (BAP1) protein expression as detected by immunohistochemistry (IHC) and homozygous deletion (HD) of the 9p21 locus as detected by fluorescence in situ hybridization (FISH) permits differentiation of MPM from reactive Mesothelial Hyperplasia (RMH). We have previously reported that detecting the loss of methylthioadenosine phosphorylase (MTAP) using IHC is a surrogate assay for 9p21 FISH. Furthermore, enhancer of zeste homolog 2 (EZH2), which encodes a component of polycomb repressor complex 2 (PRC-2), has been overexpressed in various tumors as well as MPM. In the current study, we investigated whether EZH2 IHC assay, alone or in combination with BAP1 and MTAP IHC, is useful for distinguishing MPM from RMH. Materials and methods We examined IHC expression of EZH2, BAP1, and MTAP, and 9p21 FISH in MPM (n = 38) and RMH (n = 29) and analyzed the sensitivity and specificity of each detection assay for distinguishing MPM from RMH. Results and conclusion EZH2, BAP1, and MTAP IHC, and 9p21 FISH were characterized by a 100% specificity each and 44.7%, 52.6%, 47.4%, and 65.8% sensitivities, respectively. A combination of EZH2 and BAP1 IHC, and 9p21 FISH showed the greatest sensitivity (89.5%). Using IHC alone (EZH2, BAP1, and MTAP IHC) also yielded a good sensitivity of 86.9%; this level is high enough for routine diagnostics. There were no statistically significant associations between expression of EZH2 and that of other markers (BAP1 and MTAP IHC) or 9p21 HD. However, a high expression level of EZH2 was significantly associated with short survival (P = 0.025). In conclusion, adding a high expression level of EZH2 to a combination of BAP1 and MTAP loss, all detected by IHC, demonstrated useful for discriminating MPM from RMH.

  • a combination of mtap and bap1 immunohistochemistry in pleural effusion cytology for the diagnosis of mesothelioma
    Cancer Cytopathology, 2018
    Co-Authors: Yoshiaki Kinoshita, Tohru Tsujimura, Tomoyuki Hida, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Makoto Hamasaki, Kenzo Hiroshima, Kazuki Nabeshima
    Abstract:

    BACKGROUND Homozygous deletion of 9p21 detected by fluorescence in situ hybridization (FISH) and loss of BRCA1-associated protein 1 (BAP1) expression detected by immunohistochemistry (IHC) are useful for the differentiation between malignant pleural mesothelioma (MPM) and reactive Mesothelial Hyperplasia. The authors previously described that IHC expression of the protein product of the methylthioadenosine phosphorylase (MTAP) gene, which is localized in the 9p21 chromosomal region, was correlated with the deletion status of 9p21 FISH in MPM tissues. In the current study, the authors investigated whether a combination of MTAP and BAP1 IHC could distinguish MPM from reactive Mesothelial cells (RMC) in cell blocks obtained from pleural effusions. METHODS The authors examined IHC expression of MTAP and BAP1 in cell blocks obtained from pleural effusions of 45 cases of MPM and 21 cases of reactive Mesothelial Hyperplasia. Furthermore, IHC expression of MTAP was compared with the deletion status of 9p21 FISH. RESULTS MTAP and BAP1 IHC differentiated MPM from RMC with 100% specificity for both and sensitivities of 42.2% and 60.0%, respectively. The combination of MTAP and BAP1 IHC yielded a sensitivity of 77.8%, which was higher than that of BAP1 IHC alone or 9p21 FISH alone (62.2%). Moreover, a high degree of concordance was observed between the results of MTAP IHC and 9p21 FISH in cell blocks. CONCLUSIONS A combination of MTAP and BAP1 IHC in cell blocks from pleural effusions appears to be a reliable and useful method for differentiating MPM cells from RMC and can be used in the routine diagnosis of MPM. Cancer Cytopathol 2017. © 2017 American Cancer Society.

  • immunohistochemical detection of mtap and bap1 protein loss for mesothelioma diagnosis comparison with 9p21 fish and bap1 immunohistochemistry
    Lung Cancer, 2017
    Co-Authors: Tatsuro Okamoto, Tohru Tsujimura, Tomoyuki Hida, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Makoto Hamasaki, Akinori Iwasaki, Yoshinao Oda
    Abstract:

    Abstract Objectives Differentiating malignant pleural mesothelioma (MPM) from reactive Mesothelial Hyperplasia (RMH) is still challenging. Detection of homozygous deletion (HD) of 9p21 region including p16 INK4A ( p16 ) by fluorescence in situ hybridization (FISH) and immunohistochemical detection of loss of BRCA1 associated protein 1 (BAP1), are reliable markers for MPM diagnosis. However, not all laboratories are equipped to perform 9p21 FISH; immunohistochemistry (IHC) is a more common and feasible technique. Thus, we sought to develop a IHC-based method that could predict the deletion of p16 in MPM in concordance with 9p21 FISH. Materials and methods We examined the expression of the 9p21.3-related proteins (p14, p15, p16, and methylthioadenosine phosphorylase (MTAP)) and BAP1 using IHC in 51 MPM and 25 RMH cases, and assessed their correlation with HD of p16 detected by FISH. The diagnostic usefulness of IHC of the 9p21.3-related proteins and BAP1 and their combinations was assessed using the cut-off values set by receiver operating characteristic (ROC) analysis. Results Among the 9p21.3-related proteins, MTAP IHC findings showed best concordance with 9p21 FISH results (kappa coefficient of 0.69) and a specificity of 100%. We also examined the combinations of MTAP IHC with the other products. The loss of p16 and MTAP had better concordance (kappa coefficient of 0.71), although lower specificity (85%). For differentiating MPM from RMH, only MTAP showed 100% specificity among the 9p21.3-related proteins, as did BAP1 IHC and 9p21 FISH. Among BAP1 combinations, only that of BAP1 with MTAP showed 100% specificity. Its sensitivity was 76.5%, which was lower than BAP1 IHC and 9p21 FISH combination (84.3%), but higher than BAP1 IHC alone (60.8%) or 9p21 FISH alone (60.8%). Conclusions A combination of MTAP or BAP1 loss detected by IHC can likely detect MPM with good sensitivity and 100% specificity, and serve as useful ancillary IHC for discriminating MPM from RMH.

  • bap1 immunohistochemistry and p16 fish results in combination provide higher confidence in malignant pleural mesothelioma diagnosis roc analysis of the two tests
    Pathology International, 2016
    Co-Authors: Tomoyuki Hida, Tatsuro Okamoto, Tohru Tsujimura, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Makoto Hamasaki, Akinori Iwasaki, Hiroshi Honda, Kazuki Nabeshima
    Abstract:

    Differentiation of malignant pleural mesothelioma (MPM) from benign Mesothelial proliferation remains problematic. Loss of nuclear staining of BRCA1-associated protein 1 (BAP1; detected using immunohistochemistry (IHC)) and homozygous deletion (HD) of p16 (detected using fluorescence in situ hybridization (FISH)) are useful for differentiation of MPM from reactive Mesothelial Hyperplasia (RMH), but the correlation between BAP1 expression loss and p16 HD has not been fully described. We performed BAP1 IHC and p16-specific FISH for 40 MPM and 20 RMH cases, and measured proportions of cells showing BAP1 expression and p16 HD for each case. The diagnostic accuracy for MPM and the cut-off values of the two methods were assessed using receiver operating characteristic (ROC) analysis. BAP1 expression loss, p16 HD and coexistence of both were present in 27 (67.5 %), 27 (67.5 %) and 17 (42.5 %) MPM cases, respectively. Three MPM cases (7.5 %) and all 20 RMH cases had neither BAP1 loss nor p16 HD. There was no correlation between the results of the two methods. Their combination showed higher sensitivity (92.5 %, 37/40) and estimated probability than BAP1 IHC and p16-specific FISH used alone. BAP1 IHC and p16-specific FISH have independent diagnostic value, and have increased reliability when used in combination, for MPM diagnosis.

  • deletion status of p16 in effusion smear preparation correlates with that of underlying malignant pleural mesothelioma tissue
    Cancer Science, 2015
    Co-Authors: Tohru Tsujimura, Tomoyuki Hida, Kunimitsu Kawahara, Shinji Matsumoto, Makoto Hamasaki, Kenzo Hiroshima, Toshiaki Kamei, Kenichi Taguchi, Akinori Iwasaki
    Abstract:

    Differentiating malignant pleural mesothelioma (MPM) cells morphologically from reactive Mesothelial Hyperplasia cells is problematic. Homozygous deletion (HD) of p16 (CDKN2A), detected by FISH, is a good marker of malignancy and is useful to differentiate between these cells. However, the correlation between the p16 status of effusion smears and that of the underlying MPM tissues has not been investigated. We used p16-specific FISH to investigate 20 cases of MPM from which both effusion cytologic smears and histologic specimens were available. In five cases, histologic specimens included both an invasive component and surface Mesothelial proliferation. In 14 cases (70%), MPM cells in both tissue sections and effusion smears were p16 HD-positive. Conversely, MPM cells in the remaining six tumors (30%) were p16 HD-negative in both tissue sections and effusion smears. For all five MPM cases with surface Mesothelial proliferations and invasive components, the effusion smears, surface Mesothelial proliferations, and invasive MPM components all displayed p16 deletion. Moreover, the extent to which p16 was deleted in smears highly correlated with the extent of p16 deletion in tissues. The p16 deletion percentages were also similar among smears, tissue surface proliferations, and invasive components. In cases with clinical and radiologic evidence of a diffuse pleural tumor, detection of p16 deletion in cytologic smear samples may permit MPM diagnosis without additional tissue examination. However, the absence of p16 deletion in cytologic smear samples does not preclude MPM.

A Ferrari - One of the best experts on this subject based on the ideXlab platform.

Makoto Hamasaki - One of the best experts on this subject based on the ideXlab platform.

  • highly expressed ezh2 in combination with bap1 and mtap loss as detected by immunohistochemistry is useful for differentiating malignant pleural mesothelioma from reactive Mesothelial Hyperplasia
    Lung Cancer, 2019
    Co-Authors: Masayo Yoshimura, Tomoyuki Hida, Shinji Matsumoto, Makoto Hamasaki, Akinori Iwasaki, Yoshiaki Kinoshita, Yoshinao Oda, Kazuki Nabeshima
    Abstract:

    Abstract Objective Malignant pleural mesothelioma (MPM) is an aggressive neoplasm with poor prognosis. Loss of BRCA-associated protein 1 (BAP1) protein expression as detected by immunohistochemistry (IHC) and homozygous deletion (HD) of the 9p21 locus as detected by fluorescence in situ hybridization (FISH) permits differentiation of MPM from reactive Mesothelial Hyperplasia (RMH). We have previously reported that detecting the loss of methylthioadenosine phosphorylase (MTAP) using IHC is a surrogate assay for 9p21 FISH. Furthermore, enhancer of zeste homolog 2 (EZH2), which encodes a component of polycomb repressor complex 2 (PRC-2), has been overexpressed in various tumors as well as MPM. In the current study, we investigated whether EZH2 IHC assay, alone or in combination with BAP1 and MTAP IHC, is useful for distinguishing MPM from RMH. Materials and methods We examined IHC expression of EZH2, BAP1, and MTAP, and 9p21 FISH in MPM (n = 38) and RMH (n = 29) and analyzed the sensitivity and specificity of each detection assay for distinguishing MPM from RMH. Results and conclusion EZH2, BAP1, and MTAP IHC, and 9p21 FISH were characterized by a 100% specificity each and 44.7%, 52.6%, 47.4%, and 65.8% sensitivities, respectively. A combination of EZH2 and BAP1 IHC, and 9p21 FISH showed the greatest sensitivity (89.5%). Using IHC alone (EZH2, BAP1, and MTAP IHC) also yielded a good sensitivity of 86.9%; this level is high enough for routine diagnostics. There were no statistically significant associations between expression of EZH2 and that of other markers (BAP1 and MTAP IHC) or 9p21 HD. However, a high expression level of EZH2 was significantly associated with short survival (P = 0.025). In conclusion, adding a high expression level of EZH2 to a combination of BAP1 and MTAP loss, all detected by IHC, demonstrated useful for discriminating MPM from RMH.

  • a combination of mtap and bap1 immunohistochemistry in pleural effusion cytology for the diagnosis of mesothelioma
    Cancer Cytopathology, 2018
    Co-Authors: Yoshiaki Kinoshita, Tohru Tsujimura, Tomoyuki Hida, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Makoto Hamasaki, Kenzo Hiroshima, Kazuki Nabeshima
    Abstract:

    BACKGROUND Homozygous deletion of 9p21 detected by fluorescence in situ hybridization (FISH) and loss of BRCA1-associated protein 1 (BAP1) expression detected by immunohistochemistry (IHC) are useful for the differentiation between malignant pleural mesothelioma (MPM) and reactive Mesothelial Hyperplasia. The authors previously described that IHC expression of the protein product of the methylthioadenosine phosphorylase (MTAP) gene, which is localized in the 9p21 chromosomal region, was correlated with the deletion status of 9p21 FISH in MPM tissues. In the current study, the authors investigated whether a combination of MTAP and BAP1 IHC could distinguish MPM from reactive Mesothelial cells (RMC) in cell blocks obtained from pleural effusions. METHODS The authors examined IHC expression of MTAP and BAP1 in cell blocks obtained from pleural effusions of 45 cases of MPM and 21 cases of reactive Mesothelial Hyperplasia. Furthermore, IHC expression of MTAP was compared with the deletion status of 9p21 FISH. RESULTS MTAP and BAP1 IHC differentiated MPM from RMC with 100% specificity for both and sensitivities of 42.2% and 60.0%, respectively. The combination of MTAP and BAP1 IHC yielded a sensitivity of 77.8%, which was higher than that of BAP1 IHC alone or 9p21 FISH alone (62.2%). Moreover, a high degree of concordance was observed between the results of MTAP IHC and 9p21 FISH in cell blocks. CONCLUSIONS A combination of MTAP and BAP1 IHC in cell blocks from pleural effusions appears to be a reliable and useful method for differentiating MPM cells from RMC and can be used in the routine diagnosis of MPM. Cancer Cytopathol 2017. © 2017 American Cancer Society.

  • immunohistochemical detection of mtap and bap1 protein loss for mesothelioma diagnosis comparison with 9p21 fish and bap1 immunohistochemistry
    Lung Cancer, 2017
    Co-Authors: Tatsuro Okamoto, Tohru Tsujimura, Tomoyuki Hida, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Makoto Hamasaki, Akinori Iwasaki, Yoshinao Oda
    Abstract:

    Abstract Objectives Differentiating malignant pleural mesothelioma (MPM) from reactive Mesothelial Hyperplasia (RMH) is still challenging. Detection of homozygous deletion (HD) of 9p21 region including p16 INK4A ( p16 ) by fluorescence in situ hybridization (FISH) and immunohistochemical detection of loss of BRCA1 associated protein 1 (BAP1), are reliable markers for MPM diagnosis. However, not all laboratories are equipped to perform 9p21 FISH; immunohistochemistry (IHC) is a more common and feasible technique. Thus, we sought to develop a IHC-based method that could predict the deletion of p16 in MPM in concordance with 9p21 FISH. Materials and methods We examined the expression of the 9p21.3-related proteins (p14, p15, p16, and methylthioadenosine phosphorylase (MTAP)) and BAP1 using IHC in 51 MPM and 25 RMH cases, and assessed their correlation with HD of p16 detected by FISH. The diagnostic usefulness of IHC of the 9p21.3-related proteins and BAP1 and their combinations was assessed using the cut-off values set by receiver operating characteristic (ROC) analysis. Results Among the 9p21.3-related proteins, MTAP IHC findings showed best concordance with 9p21 FISH results (kappa coefficient of 0.69) and a specificity of 100%. We also examined the combinations of MTAP IHC with the other products. The loss of p16 and MTAP had better concordance (kappa coefficient of 0.71), although lower specificity (85%). For differentiating MPM from RMH, only MTAP showed 100% specificity among the 9p21.3-related proteins, as did BAP1 IHC and 9p21 FISH. Among BAP1 combinations, only that of BAP1 with MTAP showed 100% specificity. Its sensitivity was 76.5%, which was lower than BAP1 IHC and 9p21 FISH combination (84.3%), but higher than BAP1 IHC alone (60.8%) or 9p21 FISH alone (60.8%). Conclusions A combination of MTAP or BAP1 loss detected by IHC can likely detect MPM with good sensitivity and 100% specificity, and serve as useful ancillary IHC for discriminating MPM from RMH.

  • bap1 immunohistochemistry and p16 fish results in combination provide higher confidence in malignant pleural mesothelioma diagnosis roc analysis of the two tests
    Pathology International, 2016
    Co-Authors: Tomoyuki Hida, Tatsuro Okamoto, Tohru Tsujimura, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Makoto Hamasaki, Akinori Iwasaki, Hiroshi Honda, Kazuki Nabeshima
    Abstract:

    Differentiation of malignant pleural mesothelioma (MPM) from benign Mesothelial proliferation remains problematic. Loss of nuclear staining of BRCA1-associated protein 1 (BAP1; detected using immunohistochemistry (IHC)) and homozygous deletion (HD) of p16 (detected using fluorescence in situ hybridization (FISH)) are useful for differentiation of MPM from reactive Mesothelial Hyperplasia (RMH), but the correlation between BAP1 expression loss and p16 HD has not been fully described. We performed BAP1 IHC and p16-specific FISH for 40 MPM and 20 RMH cases, and measured proportions of cells showing BAP1 expression and p16 HD for each case. The diagnostic accuracy for MPM and the cut-off values of the two methods were assessed using receiver operating characteristic (ROC) analysis. BAP1 expression loss, p16 HD and coexistence of both were present in 27 (67.5 %), 27 (67.5 %) and 17 (42.5 %) MPM cases, respectively. Three MPM cases (7.5 %) and all 20 RMH cases had neither BAP1 loss nor p16 HD. There was no correlation between the results of the two methods. Their combination showed higher sensitivity (92.5 %, 37/40) and estimated probability than BAP1 IHC and p16-specific FISH used alone. BAP1 IHC and p16-specific FISH have independent diagnostic value, and have increased reliability when used in combination, for MPM diagnosis.

  • low homozygous high heterozygous deletion status by p16 fish correlates with a better prognostic group than high homozygous deletion status in malignant pleural mesothelioma
    Lung Cancer, 2016
    Co-Authors: Makoto Hamasaki, Tohru Tsujimura, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Kenzo Hiroshima, Toshiaki Kamei, Sousei Abe, Daisuke Hamatake, Yukio Nakatani
    Abstract:

    Abstract Objectives Homozygous deletion (homo-d) of the p16 (CDKN2A) gene, as determined by fluorescence in situ hybridization (FISH), helps differentiate malignant pleural mesothelioma (MPM) from reactive Mesothelial Hyperplasia (RMH). Heterozygous deletion (hetero-d) has also been identified variably in p16 FISH. This study aimed to investigate the significance of homo-d and hetero-d of p16 in the diagnosis and prognosis of MPM. Materials and methods p16 FISH was performed in 93 MPMs and 47 RMHs. Real-time polymerase chain reaction (PCR) and methylation specific PCR (MSP) were also performed for cases in which DNA was available. Overall survival (OS) was assessed via the Kaplan-Meier method and logrank test. Results Cutoff values for homo-d and hetero-d were set at 10% and 47%, respectively, based on p16 FISH results in RMH. In MPM, 80/93 (86.0%) were homo-d positive, and 15/93 (16.1%) were hetero-d positive. No RMH was homo/hetero-d positive. In nine cases of MPM with the low homo-d ( 47%) pattern, FISH with a shorter probe caused a slight increase (from 20.1% to 26.5%) in the mean percentage of homo-d and a decrease in that of hetero-d (from 59.6% to 55.6%). Four cases in which the low homo-d/high hetero-d pattern was maintained with the shorter probe were further analyzed by real-time PCR, which separated them into a two (n = 2) or one allele deletion group (n = 2). MSP revealed no promoter methylation in the two cases with one allele deletion. The OS was significantly shorter in homo-d positive cases (n = 24) than homo-d negative cases (n = 5, p = 0.0002) in the 29 MPM cases with follow-up data. Also, low homo-d/high hetero-d cases (n = 5) had a significantly better prognosis than high homo-d (≥30%) cases (n = 17, p = 0.011). Conclusions Within p16 homo-d positive MPMs with poorer prognosis, the low homo-d/high hetero-d pattern may belong to a better prognostic subgroup in homo-d positive MPMs.

Kenzo Hiroshima - One of the best experts on this subject based on the ideXlab platform.

  • a combination of mtap and bap1 immunohistochemistry in pleural effusion cytology for the diagnosis of mesothelioma
    Cancer Cytopathology, 2018
    Co-Authors: Yoshiaki Kinoshita, Tohru Tsujimura, Tomoyuki Hida, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Makoto Hamasaki, Kenzo Hiroshima, Kazuki Nabeshima
    Abstract:

    BACKGROUND Homozygous deletion of 9p21 detected by fluorescence in situ hybridization (FISH) and loss of BRCA1-associated protein 1 (BAP1) expression detected by immunohistochemistry (IHC) are useful for the differentiation between malignant pleural mesothelioma (MPM) and reactive Mesothelial Hyperplasia. The authors previously described that IHC expression of the protein product of the methylthioadenosine phosphorylase (MTAP) gene, which is localized in the 9p21 chromosomal region, was correlated with the deletion status of 9p21 FISH in MPM tissues. In the current study, the authors investigated whether a combination of MTAP and BAP1 IHC could distinguish MPM from reactive Mesothelial cells (RMC) in cell blocks obtained from pleural effusions. METHODS The authors examined IHC expression of MTAP and BAP1 in cell blocks obtained from pleural effusions of 45 cases of MPM and 21 cases of reactive Mesothelial Hyperplasia. Furthermore, IHC expression of MTAP was compared with the deletion status of 9p21 FISH. RESULTS MTAP and BAP1 IHC differentiated MPM from RMC with 100% specificity for both and sensitivities of 42.2% and 60.0%, respectively. The combination of MTAP and BAP1 IHC yielded a sensitivity of 77.8%, which was higher than that of BAP1 IHC alone or 9p21 FISH alone (62.2%). Moreover, a high degree of concordance was observed between the results of MTAP IHC and 9p21 FISH in cell blocks. CONCLUSIONS A combination of MTAP and BAP1 IHC in cell blocks from pleural effusions appears to be a reliable and useful method for differentiating MPM cells from RMC and can be used in the routine diagnosis of MPM. Cancer Cytopathol 2017. © 2017 American Cancer Society.

  • low homozygous high heterozygous deletion status by p16 fish correlates with a better prognostic group than high homozygous deletion status in malignant pleural mesothelioma
    Lung Cancer, 2016
    Co-Authors: Makoto Hamasaki, Tohru Tsujimura, Kunimitsu Kawahara, Ayuko Sato, Shinji Matsumoto, Kenzo Hiroshima, Toshiaki Kamei, Sousei Abe, Daisuke Hamatake, Yukio Nakatani
    Abstract:

    Abstract Objectives Homozygous deletion (homo-d) of the p16 (CDKN2A) gene, as determined by fluorescence in situ hybridization (FISH), helps differentiate malignant pleural mesothelioma (MPM) from reactive Mesothelial Hyperplasia (RMH). Heterozygous deletion (hetero-d) has also been identified variably in p16 FISH. This study aimed to investigate the significance of homo-d and hetero-d of p16 in the diagnosis and prognosis of MPM. Materials and methods p16 FISH was performed in 93 MPMs and 47 RMHs. Real-time polymerase chain reaction (PCR) and methylation specific PCR (MSP) were also performed for cases in which DNA was available. Overall survival (OS) was assessed via the Kaplan-Meier method and logrank test. Results Cutoff values for homo-d and hetero-d were set at 10% and 47%, respectively, based on p16 FISH results in RMH. In MPM, 80/93 (86.0%) were homo-d positive, and 15/93 (16.1%) were hetero-d positive. No RMH was homo/hetero-d positive. In nine cases of MPM with the low homo-d ( 47%) pattern, FISH with a shorter probe caused a slight increase (from 20.1% to 26.5%) in the mean percentage of homo-d and a decrease in that of hetero-d (from 59.6% to 55.6%). Four cases in which the low homo-d/high hetero-d pattern was maintained with the shorter probe were further analyzed by real-time PCR, which separated them into a two (n = 2) or one allele deletion group (n = 2). MSP revealed no promoter methylation in the two cases with one allele deletion. The OS was significantly shorter in homo-d positive cases (n = 24) than homo-d negative cases (n = 5, p = 0.0002) in the 29 MPM cases with follow-up data. Also, low homo-d/high hetero-d cases (n = 5) had a significantly better prognosis than high homo-d (≥30%) cases (n = 17, p = 0.011). Conclusions Within p16 homo-d positive MPMs with poorer prognosis, the low homo-d/high hetero-d pattern may belong to a better prognostic subgroup in homo-d positive MPMs.

  • peritoneal malignant mesothelioma pmm and primary peritoneal serous carcinoma ppsc and reactive Mesothelial Hyperplasia rmh of the peritoneum immunohistochemical and fluorescence in situ hybridisation fish analyses
    Journal of Clinical Pathology, 2016
    Co-Authors: Toshiaki Kawai, Kunimitsu Kawahara, Susumu Tominaga, Sadayuki Hiroi, Sho Ogata, Kuniaki Nakanishi, Hiroshi Sonobe, Kenzo Hiroshima
    Abstract:

    Aims Peritoneal malignant mesothelioma (PMM) is an uncommon tumour, accounting for only 7–9% of all mesotheliomas in Japan. Differential diagnosis between PMM and primary peritoneal serous carcinoma (PPSC), a high-grade serous carcinoma, may be difficult, and separating reactive Mesothelial Hyperplasia (RMH) from PMM can be even more challenging. Methods To help differentiate PMM from PPSC and RMH, we used immunohistochemistry to examine Mesothelial-associated markers (calretinin, AE1/AE3, CK5/6, CAM5.2, D2-40, WT-1, HBME1, thrombomodulin), adenocarcinoma-associated markers (CEA, BerEP4, MOC31, ER (estrogen receptor), PgR, TTF-1, Claudin-4, Pax8), and malignant-related and benign-related markers (epithelial membrane antigen (EMA), desmin, GLUT-1, CD146 and IMP3), and FISH to examine for homozygous deletion of 9p21. We used formalin-fixed, paraffin-embedded blocks from 22 PMMs (M:F=18:4; subtypes: 16 epithelioid, 6 biphasic), 11 PPSCs and 23 RMHs. Results Seventeen of the mesotheliomas (four PMM from women) were classified as diffuse, while five were localised. Calretinin was 91% positive in PMM, but negative in PPSC (specificity, 100%). BerEP4, Claudin-4 and PAX8 were all 100% positive in PPSC (specificities, 100%, 95% and 95%, respectively, for excluding PMM). For distinguishing PMM and RMH, sensitivity for EMA in mesothelioma was 68%, while for IMP3 and GLUT-1 it was 64% and 50%, respectively, all with high specificities. FISH analysis revealed homozygous deletion of the 9p21 locus in 11/13 PMMs, but in 0/11 RMHs. Conclusions Calretinin and BerEP4 may be the best positive markers for differentiating PMM from PPSC. EMA, in combination with IMP3 and desmin, is useful, and homozygous deletion of 9p21 may be helpful, for differentiating PMM from RMH.

  • deletion status of p16 in effusion smear preparation correlates with that of underlying malignant pleural mesothelioma tissue
    Cancer Science, 2015
    Co-Authors: Tohru Tsujimura, Tomoyuki Hida, Kunimitsu Kawahara, Shinji Matsumoto, Makoto Hamasaki, Kenzo Hiroshima, Toshiaki Kamei, Kenichi Taguchi, Akinori Iwasaki
    Abstract:

    Differentiating malignant pleural mesothelioma (MPM) cells morphologically from reactive Mesothelial Hyperplasia cells is problematic. Homozygous deletion (HD) of p16 (CDKN2A), detected by FISH, is a good marker of malignancy and is useful to differentiate between these cells. However, the correlation between the p16 status of effusion smears and that of the underlying MPM tissues has not been investigated. We used p16-specific FISH to investigate 20 cases of MPM from which both effusion cytologic smears and histologic specimens were available. In five cases, histologic specimens included both an invasive component and surface Mesothelial proliferation. In 14 cases (70%), MPM cells in both tissue sections and effusion smears were p16 HD-positive. Conversely, MPM cells in the remaining six tumors (30%) were p16 HD-negative in both tissue sections and effusion smears. For all five MPM cases with surface Mesothelial proliferations and invasive components, the effusion smears, surface Mesothelial proliferations, and invasive MPM components all displayed p16 deletion. Moreover, the extent to which p16 was deleted in smears highly correlated with the extent of p16 deletion in tissues. The p16 deletion percentages were also similar among smears, tissue surface proliferations, and invasive components. In cases with clinical and radiologic evidence of a diffuse pleural tumor, detection of p16 deletion in cytologic smear samples may permit MPM diagnosis without additional tissue examination. However, the absence of p16 deletion in cytologic smear samples does not preclude MPM.

  • p16 cdkn2a fish in differentiation of diffuse malignant peritoneal mesothelioma from Mesothelial Hyperplasia and epithelial ovarian cancer
    American Journal of Clinical Pathology, 2015
    Co-Authors: Tomohiro Ito, Tohru Tsujimura, Shinji Matsumoto, Makoto Hamasaki, Kenzo Hiroshima, Toshiaki Kawai, Yoshiya Shimao, Kousuke Marutsuka, Sayaka Moriguchi, Riruke Maruyama
    Abstract:

    Objectives: It can be difficult to differentiate diffuse malignant peritoneal mesothelioma (DMPM) from reactive Mesothelial Hyperplasia (RMH) or peritoneal dissemination of gynecologic malignancies, such as epithelial ovarian cancer (EOC), which cause a large amount of ascites. Detection of the homozygous deletion of p16/CDKN2A ( p16 ) by fluorescence in situ hybridization (FISH) is an effective adjunct in the diagnosis of malignant pleural mesothelioma. The aim of this study was to investigate the ability of the p16 FISH assay to differentiate DMPM from RMH and EOC. Methods: p16 FISH was performed in 28 DMPMs (successful in 19), 30 RMHs, and 40 EOC cases. The cutoff values of p16 FISH were more than 10% for homozygous deletion and more than 40% for heterozygous deletion. Results: According to the above criteria, nine (47.4%) of 19 successful DMPM cases were homozygous deletion positive, and three (15.8%) of 19 were heterozygous deletion positive, whereas all RMH cases were negative for the p16 deletion. In all four major histologic subtypes of EOC, neither p16 homozygous nor heterozygous deletions were detected. To differentiate DMPM from RMH or EOC, the sensitivity of the p16 homozygous deletion was 32% (9/28), and the specificity was 100%. Conclusions: Our study suggests that p16 FISH analysis is useful in differentiating DMPM from RMH and EOC when homozygous deletion is detected.