In depth evaluation of the prognostic and predictive utility of PTEN immunohistochemistry in colorectal carcinomas: Performance of three antibodies with emphasis on intracellular and intratumoral heterogeneity

Emese Irma Ágoston, Tamás Micsik, Balázs Ács, Krisztina Fekete, Oszkár Hahn, Zsolt Baranyai, Kristóf Dede, György Bodoky, Attila Bursics, Janina Kulka, Tibor Krenács, B. Györffy, László Harsányi, A. Szász

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Abstract

Background: Phosphatase and tensin homolog deleted in chromosome 10 (PTEN) loss of function is frequently detected in advanced colorectal cancer. Its detection is thought to have prognostic significance and it is being considered to predict responsiveness to anti-EGFR therapy. Unfortunately, while immunohistochemical assessment of PTEN expression is widespread, it lacks standardization and the results are hardly comparable across the available publications. Methods: Retrospectively collected, formalin-fixed and paraffin-embedded colorectal tumor tissue samples from 55 patients were combined into tissue microarray (TMA) blocks. We used three different PTEN antibodies to determine the frequency, intensity and intracellular pattern of PTEN immunohistochemical labeling: Neomarkers, Dako and CellSignaling. We evaluated the aforementioned parameters in selected regions of colorectal cancers and in their lymph node metastases by using three scoring methods that take into consideration both staining frequency and intensity (H1-H3-score). We also evaluated intracellular localization. Results: The Dako and CellSignaling antibodies stained predominantly cytoplasms, while the Neomarkers antibody specifically stained cell nuclei. PTEN H-scores were significantly lower in all tumor areas as compared to the normal colonic mucosa based on staining with the DAKO and CellSignaling antibodies. Intratumoral regional differences or differences between matching tumors and metastases were not detected with any of the antibodies. Neither Dako, neither CellSignaling, nor the Neomarkers antibodies revealed a significant correlation between PTEN expression and pT, Dukes/MAC and clinical stage. KRAS status, histological grade correlated with PTEN H-scores based on staining with the Neomarkers antibody. PTEN H-scores did not correlate with MMR status. PTEN H-scores did not show any correlation with relapse-free survival based on staining with either antibody. Conclusions: While PTEN expression decreased in colorectal cancer according to two antibodies, neither of the three applied PTEN antibodies could justify significant correlation with clinicopathological data, nor had prognostic value. Thus, we might conclude that immunohistochemical PTEN investigation remains a challenge requiring more standardized evaluation on larger number of cases to clarify its utility as a prognostic and predictive tool in CRC. The standardization of immunohistochemical method is key in the evaluation process, which is further discussed.

Original languageEnglish
Article number61
JournalDiagnostic Pathology
Volume11
Issue number1
DOIs
Publication statusPublished - 2016

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Colorectal Neoplasms
Immunohistochemistry
Antibodies
Staining and Labeling
Neoplasm Metastasis
Chromosomes, Human, Pair 10
Cell Nucleus
Phosphoric Monoester Hydrolases
Paraffin
Formaldehyde
Publications
Neoplasms
Cytoplasm
Mucous Membrane
Research Design
Lymph Nodes
Recurrence
Survival

Keywords

  • Colorectal cancer
  • Immunohistochemistry
  • Intratumoral heterogeneity
  • Predictive marker
  • Prognostic marker
  • Protein expression
  • PTEN

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

Cite this

In depth evaluation of the prognostic and predictive utility of PTEN immunohistochemistry in colorectal carcinomas : Performance of three antibodies with emphasis on intracellular and intratumoral heterogeneity. / Ágoston, Emese Irma; Micsik, Tamás; Ács, Balázs; Fekete, Krisztina; Hahn, Oszkár; Baranyai, Zsolt; Dede, Kristóf; Bodoky, György; Bursics, Attila; Kulka, Janina; Krenács, Tibor; Györffy, B.; Harsányi, László; Szász, A.

In: Diagnostic Pathology, Vol. 11, No. 1, 61, 2016.

Research output: Contribution to journalArticle

Ágoston, Emese Irma ; Micsik, Tamás ; Ács, Balázs ; Fekete, Krisztina ; Hahn, Oszkár ; Baranyai, Zsolt ; Dede, Kristóf ; Bodoky, György ; Bursics, Attila ; Kulka, Janina ; Krenács, Tibor ; Györffy, B. ; Harsányi, László ; Szász, A. / In depth evaluation of the prognostic and predictive utility of PTEN immunohistochemistry in colorectal carcinomas : Performance of three antibodies with emphasis on intracellular and intratumoral heterogeneity. In: Diagnostic Pathology. 2016 ; Vol. 11, No. 1.
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abstract = "Background: Phosphatase and tensin homolog deleted in chromosome 10 (PTEN) loss of function is frequently detected in advanced colorectal cancer. Its detection is thought to have prognostic significance and it is being considered to predict responsiveness to anti-EGFR therapy. Unfortunately, while immunohistochemical assessment of PTEN expression is widespread, it lacks standardization and the results are hardly comparable across the available publications. Methods: Retrospectively collected, formalin-fixed and paraffin-embedded colorectal tumor tissue samples from 55 patients were combined into tissue microarray (TMA) blocks. We used three different PTEN antibodies to determine the frequency, intensity and intracellular pattern of PTEN immunohistochemical labeling: Neomarkers, Dako and CellSignaling. We evaluated the aforementioned parameters in selected regions of colorectal cancers and in their lymph node metastases by using three scoring methods that take into consideration both staining frequency and intensity (H1-H3-score). We also evaluated intracellular localization. Results: The Dako and CellSignaling antibodies stained predominantly cytoplasms, while the Neomarkers antibody specifically stained cell nuclei. PTEN H-scores were significantly lower in all tumor areas as compared to the normal colonic mucosa based on staining with the DAKO and CellSignaling antibodies. Intratumoral regional differences or differences between matching tumors and metastases were not detected with any of the antibodies. Neither Dako, neither CellSignaling, nor the Neomarkers antibodies revealed a significant correlation between PTEN expression and pT, Dukes/MAC and clinical stage. KRAS status, histological grade correlated with PTEN H-scores based on staining with the Neomarkers antibody. PTEN H-scores did not correlate with MMR status. PTEN H-scores did not show any correlation with relapse-free survival based on staining with either antibody. Conclusions: While PTEN expression decreased in colorectal cancer according to two antibodies, neither of the three applied PTEN antibodies could justify significant correlation with clinicopathological data, nor had prognostic value. Thus, we might conclude that immunohistochemical PTEN investigation remains a challenge requiring more standardized evaluation on larger number of cases to clarify its utility as a prognostic and predictive tool in CRC. The standardization of immunohistochemical method is key in the evaluation process, which is further discussed.",
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T1 - In depth evaluation of the prognostic and predictive utility of PTEN immunohistochemistry in colorectal carcinomas

T2 - Performance of three antibodies with emphasis on intracellular and intratumoral heterogeneity

AU - Ágoston, Emese Irma

AU - Micsik, Tamás

AU - Ács, Balázs

AU - Fekete, Krisztina

AU - Hahn, Oszkár

AU - Baranyai, Zsolt

AU - Dede, Kristóf

AU - Bodoky, György

AU - Bursics, Attila

AU - Kulka, Janina

AU - Krenács, Tibor

AU - Györffy, B.

AU - Harsányi, László

AU - Szász, A.

PY - 2016

Y1 - 2016

N2 - Background: Phosphatase and tensin homolog deleted in chromosome 10 (PTEN) loss of function is frequently detected in advanced colorectal cancer. Its detection is thought to have prognostic significance and it is being considered to predict responsiveness to anti-EGFR therapy. Unfortunately, while immunohistochemical assessment of PTEN expression is widespread, it lacks standardization and the results are hardly comparable across the available publications. Methods: Retrospectively collected, formalin-fixed and paraffin-embedded colorectal tumor tissue samples from 55 patients were combined into tissue microarray (TMA) blocks. We used three different PTEN antibodies to determine the frequency, intensity and intracellular pattern of PTEN immunohistochemical labeling: Neomarkers, Dako and CellSignaling. We evaluated the aforementioned parameters in selected regions of colorectal cancers and in their lymph node metastases by using three scoring methods that take into consideration both staining frequency and intensity (H1-H3-score). We also evaluated intracellular localization. Results: The Dako and CellSignaling antibodies stained predominantly cytoplasms, while the Neomarkers antibody specifically stained cell nuclei. PTEN H-scores were significantly lower in all tumor areas as compared to the normal colonic mucosa based on staining with the DAKO and CellSignaling antibodies. Intratumoral regional differences or differences between matching tumors and metastases were not detected with any of the antibodies. Neither Dako, neither CellSignaling, nor the Neomarkers antibodies revealed a significant correlation between PTEN expression and pT, Dukes/MAC and clinical stage. KRAS status, histological grade correlated with PTEN H-scores based on staining with the Neomarkers antibody. PTEN H-scores did not correlate with MMR status. PTEN H-scores did not show any correlation with relapse-free survival based on staining with either antibody. Conclusions: While PTEN expression decreased in colorectal cancer according to two antibodies, neither of the three applied PTEN antibodies could justify significant correlation with clinicopathological data, nor had prognostic value. Thus, we might conclude that immunohistochemical PTEN investigation remains a challenge requiring more standardized evaluation on larger number of cases to clarify its utility as a prognostic and predictive tool in CRC. The standardization of immunohistochemical method is key in the evaluation process, which is further discussed.

AB - Background: Phosphatase and tensin homolog deleted in chromosome 10 (PTEN) loss of function is frequently detected in advanced colorectal cancer. Its detection is thought to have prognostic significance and it is being considered to predict responsiveness to anti-EGFR therapy. Unfortunately, while immunohistochemical assessment of PTEN expression is widespread, it lacks standardization and the results are hardly comparable across the available publications. Methods: Retrospectively collected, formalin-fixed and paraffin-embedded colorectal tumor tissue samples from 55 patients were combined into tissue microarray (TMA) blocks. We used three different PTEN antibodies to determine the frequency, intensity and intracellular pattern of PTEN immunohistochemical labeling: Neomarkers, Dako and CellSignaling. We evaluated the aforementioned parameters in selected regions of colorectal cancers and in their lymph node metastases by using three scoring methods that take into consideration both staining frequency and intensity (H1-H3-score). We also evaluated intracellular localization. Results: The Dako and CellSignaling antibodies stained predominantly cytoplasms, while the Neomarkers antibody specifically stained cell nuclei. PTEN H-scores were significantly lower in all tumor areas as compared to the normal colonic mucosa based on staining with the DAKO and CellSignaling antibodies. Intratumoral regional differences or differences between matching tumors and metastases were not detected with any of the antibodies. Neither Dako, neither CellSignaling, nor the Neomarkers antibodies revealed a significant correlation between PTEN expression and pT, Dukes/MAC and clinical stage. KRAS status, histological grade correlated with PTEN H-scores based on staining with the Neomarkers antibody. PTEN H-scores did not correlate with MMR status. PTEN H-scores did not show any correlation with relapse-free survival based on staining with either antibody. Conclusions: While PTEN expression decreased in colorectal cancer according to two antibodies, neither of the three applied PTEN antibodies could justify significant correlation with clinicopathological data, nor had prognostic value. Thus, we might conclude that immunohistochemical PTEN investigation remains a challenge requiring more standardized evaluation on larger number of cases to clarify its utility as a prognostic and predictive tool in CRC. The standardization of immunohistochemical method is key in the evaluation process, which is further discussed.

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KW - Immunohistochemistry

KW - Intratumoral heterogeneity

KW - Predictive marker

KW - Prognostic marker

KW - Protein expression

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