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Reproducibility and Prognostic Potential of Ki-67 Proliferation Index when Comparing Digital-Image Analysis with Standard Semi-Quantitative Evaluation in Breast Cancer

  • Original Article
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Pathology & Oncology Research

Abstract

In this study, the reproducibility of Ki-67 proliferation index (KIPI) was investigated by comparing the semi-quantitative (SQ) results of three assessors with those of digital image-analysis (DIA) methods. The prognostic significance of the two approaches was also correlated with clinical outcome. Tissue microarrays of duplicate 2 mm cores were constructed from representative areas of formalin-fixed and paraffin-embedded tumor blocks of 347 breast cancer patients. SQ evaluation of Ki-67 (MIB1 clone) immunostained slides was performed independently by three pathologists. DIA was completed using a fully automated histological pattern and cell recognition module for KIPI detection (DIA-1) and an adjustable module (DIA-2) with the possibility of manual corrections. To compare SQ and DIA evaluations intra-class correlation (ICC) and concordance correlation coefficients (CCC) were determined. The three SQ evaluations demonstrated a remarkable ICC (0.853). Significant difference and poor concordance occurred between SQ-1 and SQ-2 as well as between SQ-1 and SQ-3 (p ≤ 0.001, CCC ≤ 0.827 for both comparisons). Thus, the reference KIPI value (SQ-RV) was generated from the mean values of SQ-2 and SQ-3. SQ-RV and DIA-2 results showed substantial concordance (CCC = 0.963, at p = 0.754), while SQ-RV and DIA-1 values differed (p ≤ 0.001) at only moderate concordance (CCC = 0.906). In multivariate analysis, lymph node status and SQ-2 assessment were significantly associated with clinical outcome (p ≤ 0.012 for both comparisons). Our results confirm that KIPI is a significant prognostic marker in breast cancer, which can be can be reliably reproduced by using an adjustable DIA-2 image analysis module.

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Acknowledgements

Balazs Acs was supported by the Hungarian Talent Program, National Young Talent Award 2016 (NTP-NFTÖ-16-0496). Anna-Maria Tokes was supported by the Hungarian Society of Medical Oncology (MKOT) 2014-2016, and A. Marcell Szasz was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences.

We thank Gabor Kiszler Ph.D. for his innovative technical and IT support and Zsuzsanna Bodor M.D. for her excellent remarks on this study. We acknowledge Professor Zoltán Prohászka for his constructive comments on statistical analyses. We also appreciate Rita Keszthelyi’s efforts made on implementation of immunohistochemical reactions.

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Authors and Affiliations

  1. 2nd Department of Pathology, Semmelweis University, 93 Üllői út, Budapest, 1091, Hungary

    Balázs Ács, Lilla Madaras, Kristóf Attila Kovács, Janina Kulka & Attila Marcell Szász

  2. 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, 26 Üllői út, Budapest, 1085, Hungary

    Tamás Micsik

  3. MTA-SE Tumor Progression Research Group, 2nd Department of Pathology, Semmelweis University, 93 Üllői út, Budapest, 1091, Hungary

    Anna-Mária Tőkés

  4. MTA-TTK Lendület Cancer Biomarker Research Group, 2 Magyar tudósok körútja, Budapest, 1117, Hungary

    Balázs Győrffy & Attila Marcell Szász

  5. 2nd Department of Pediatrics, Semmelweis University, Tűzoltó u. 7-9, Budapest, Hungary

    Balázs Győrffy

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  1. Balázs Ács
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Contributions

BÁ carried out the conception and design of the study, performed digital-image analysis and statistical analyses, wrote the manuscript. LM evaluated Ki-67 IHC and revised the manuscript. KAK participated in the evaluation of Ki-67 IHC and drafted the manuscript. TM participated in digital-image analysis and helped to revise the manuscript. AMT reviewed follow-up data and drafted the manuscript. BG helped to interpret data and wrote the manuscript. JK designed and coordinated the study and revised the manuscript. AMSZ conceived of the study, and participated in its design and coordination, also evaluated Ki-67 IHC reactions, drafted the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Attila Marcell Szász.

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Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

The study was approved by the Institutional Review Board of Semmelweis University (IKEB, #7–1/2008). This article does not contain any studies with animals performed by any of the authors.

Informed Consent

The Institutional Review Board of Semmelweis University was consulted for patient consent requirement, written approval was unnecessary due to the nature of the study, namely routine material was utilized for the evaluation of Ki-67 without any identification of the patients and their personal data. The Helsinki Declaration of 1964 was followed in any such survey.

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Ács, B., Madaras, L., Kovács, K.A. et al. Reproducibility and Prognostic Potential of Ki-67 Proliferation Index when Comparing Digital-Image Analysis with Standard Semi-Quantitative Evaluation in Breast Cancer. Pathol. Oncol. Res. 24, 115–127 (2018). https://doi.org/10.1007/s12253-017-0220-8

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  • DOI: https://doi.org/10.1007/s12253-017-0220-8

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