Skip to main content
Top
Published in: Diagnostic Pathology 1/2018

Open Access 01-12-2018 | Research

Evaluation of the proliferation marker Ki-67 in gliomas: Interobserver variability and digital quantification

Authors: Ljudmilla A. G. Nielsen, Julie A. Bangsø, Kim H. Lindahl, Rikke H. Dahlrot, Jacob v. B. Hjelmborg, Steinbjørn Hansen, Bjarne W. Kristensen

Published in: Diagnostic Pathology | Issue 1/2018

Login to get access

Abstract

Background

The Ki-67 Labelling Index (LI) is used as an ancillary tool in glioma diagnostics. Interobserver variability has been reported and no precise guidelines are available. Nor is it known whether novel digital approaches would be an advantage. Our aim was to evaluate the inter- and intraobserver variability of the Ki-67 LI between two pathologists and between pathologists and digital quantification both in whole tumour slides and in hot spots using narrow but diagnostically relevant intervals.

Methods

In samples of 235 low and high grade gliomas, two pathologists (A and B) estimated the Ki-67 LI (5–10% intervals) for whole tumour slides and for hot spots. In 20 of the cases intraobserver variability was evaluated. For digital quantification (C) slides were scanned with subsequent systematic random sampling of viable tumour areas. A software classifier trained to identify positive and negative nuclei calculated the Ki-67 LI. The interobserver agreements were evaluated using kappa (κ) statistics.

Results

The observed proportions of agreement and κ values for Ki-67 LI for whole tumour slides were: A/B: 46% (κ = 0.32); A/C: 37% (κ = 0.26); B/C: 37% (κ = 0.26). For hot spots equivalent values were: A/B: 14% (κ = 0.04); A/C: 18% (κ = 0.09); B/C: 31% (κ = 0.21).

Conclusions

Interobserver variability was pronounced between pathologists and for pathologists versus digital quantification when attempting to estimate a precise value of the Ki-67 LI. Ki-67 LI should therefore be used with caution and should not be over interpreted in the grading of gliomas. Digital quantification of Ki-67 LI in gliomas was feasible, but intra- and interlaboratory robustness need to be determined.
Appendix
Available only for authorised users
Literature
1.
go back to reference Louis DN, Ohgaki H, Wiestler OD, Cavenee WK (Eds.). WHO classification of Tumours of the central nervous system. 4th Ed. revised. Lyon: IARC; 2016. Louis DN, Ohgaki H, Wiestler OD, Cavenee WK (Eds.). WHO classification of Tumours of the central nervous system. 4th Ed. revised. Lyon: IARC; 2016.
2.
go back to reference Mittler MA, Walters BC, Stopa EG. Observer reliability in histological grading of astrocytoma stereotactic biopsies. J Neurosurg. 1996;85(6):1091–4.CrossRefPubMed Mittler MA, Walters BC, Stopa EG. Observer reliability in histological grading of astrocytoma stereotactic biopsies. J Neurosurg. 1996;85(6):1091–4.CrossRefPubMed
3.
go back to reference Torp SH. Diagnostic and Prognostic role of Ki67 immunostaining in human astrocytomas using four different antibodies. Clin Neuropathol. 2002;21(6):252–7.PubMed Torp SH. Diagnostic and Prognostic role of Ki67 immunostaining in human astrocytomas using four different antibodies. Clin Neuropathol. 2002;21(6):252–7.PubMed
4.
go back to reference Scholzen T, Gerdes J. The Ki-67 protein: from the known and the unknown. J Cell Physiol. 2000;182:311–22.CrossRefPubMed Scholzen T, Gerdes J. The Ki-67 protein: from the known and the unknown. J Cell Physiol. 2000;182:311–22.CrossRefPubMed
5.
go back to reference Johannessen AL, Torp SH. The clinical value of Ki-67/MIB-1 labeling index in human astrocytomas. Pathol Oncol Res. 2006;12(3):143–7.CrossRefPubMed Johannessen AL, Torp SH. The clinical value of Ki-67/MIB-1 labeling index in human astrocytomas. Pathol Oncol Res. 2006;12(3):143–7.CrossRefPubMed
6.
go back to reference Prayson RA. The utility of MIB-1/Ki-67 immunostaining in the evaluation of central nervous system neoplasms. Adv Anat Pathol. 2005;12(3):144–8.CrossRefPubMed Prayson RA. The utility of MIB-1/Ki-67 immunostaining in the evaluation of central nervous system neoplasms. Adv Anat Pathol. 2005;12(3):144–8.CrossRefPubMed
7.
go back to reference Rathi KR, Radotra BD, Khosla VK. Proliferative index in astrocytic tumours. Indian J Pathol Microbiol. 2007;50(4):754–8.PubMed Rathi KR, Radotra BD, Khosla VK. Proliferative index in astrocytic tumours. Indian J Pathol Microbiol. 2007;50(4):754–8.PubMed
8.
go back to reference Grzybicki DM, Liu Y, Moore SA, Brown HG, Silverman JF, D'Amico F, Raab SS. Interobserver variability associated with the MIB-1 labeling index: high levels suggest limited prognostic usefulness for patients with primary brain tumors. Cancer. 2001;92(10):2720–6.CrossRefPubMed Grzybicki DM, Liu Y, Moore SA, Brown HG, Silverman JF, D'Amico F, Raab SS. Interobserver variability associated with the MIB-1 labeling index: high levels suggest limited prognostic usefulness for patients with primary brain tumors. Cancer. 2001;92(10):2720–6.CrossRefPubMed
9.
go back to reference Berghoff AS, Stefanits H, Woehrer A, Heinzl H, Preusser M, Hainfellner JA. Clinical neuropathology practice guide 3-2013: levels of evidence and clinical utility of prognostic and predictive candidate brain tumor biomarkers. Clin Neuropathol. 2013;32(3):148–58.CrossRefPubMedPubMedCentral Berghoff AS, Stefanits H, Woehrer A, Heinzl H, Preusser M, Hainfellner JA. Clinical neuropathology practice guide 3-2013: levels of evidence and clinical utility of prognostic and predictive candidate brain tumor biomarkers. Clin Neuropathol. 2013;32(3):148–58.CrossRefPubMedPubMedCentral
10.
go back to reference Hsu CY, Ho DM, Yang CF, Chiang H. Interobserver reproducibility of MIB-1 labeling index in astrocytic tumors using different counting methods. Mod Pathol. 2003;16(9):951–7.CrossRefPubMed Hsu CY, Ho DM, Yang CF, Chiang H. Interobserver reproducibility of MIB-1 labeling index in astrocytic tumors using different counting methods. Mod Pathol. 2003;16(9):951–7.CrossRefPubMed
11.
go back to reference Grala B, Markiewicz T, Kozlowski W, Osowski S, Slodkowska J, Papierz W. New automated image analysis method for the assessment of Ki-67 labeling index in meningiomas. Folia Histochem Cytobiol. 2009;47(4):587–92.PubMed Grala B, Markiewicz T, Kozlowski W, Osowski S, Slodkowska J, Papierz W. New automated image analysis method for the assessment of Ki-67 labeling index in meningiomas. Folia Histochem Cytobiol. 2009;47(4):587–92.PubMed
12.
go back to reference Fisher BJ, Naumova E, Leighton CC, Naumov GN, Kerklviet N, Fortin D, Macdonald DR, Cairncross JG, Bauman GS, Stitt L. Ki-67: a prognostic factor for low-grade glioma? Int J Radiat Oncol Biol Phys. 2002;52(4):996–1001.CrossRefPubMed Fisher BJ, Naumova E, Leighton CC, Naumov GN, Kerklviet N, Fortin D, Macdonald DR, Cairncross JG, Bauman GS, Stitt L. Ki-67: a prognostic factor for low-grade glioma? Int J Radiat Oncol Biol Phys. 2002;52(4):996–1001.CrossRefPubMed
13.
go back to reference Amin MR, Kurosaki M, Watanabe T, Tanaka S, Hori T. A comparative study of MIB-1 staining indices of gliomas measured by NIH image analysis program and conventional manual cell counting method. Neurol Res. 2000;22(5):495–500.CrossRefPubMed Amin MR, Kurosaki M, Watanabe T, Tanaka S, Hori T. A comparative study of MIB-1 staining indices of gliomas measured by NIH image analysis program and conventional manual cell counting method. Neurol Res. 2000;22(5):495–500.CrossRefPubMed
14.
go back to reference Vankalakunti M, Vasishta RK, Das Radotra B, Khosla VK. MIB-1 immunolabeling: a valuable marker in prediction of benign recurring meningiomas. Neuropathology. 2007;27(5):407–12. Vankalakunti M, Vasishta RK, Das Radotra B, Khosla VK. MIB-1 immunolabeling: a valuable marker in prediction of benign recurring meningiomas. Neuropathology. 2007;27(5):407–12.
15.
go back to reference Preusser M, Hoeftberger R, Woehrer A, Gelpi E, Kouwenhoven M, Kros JM, Sanson M, Idbaih A, Brandes AA, Heinzl H, Gorlia T, Hainfellner JA, van den bent M. Prognostic value of Ki67 index in anaplastic oligodendroglial tumours--a translational study of the European Organization for Research and Treatment of Cancer brain tumor group. Histopathology. 2012;60(6):885–94. Preusser M, Hoeftberger R, Woehrer A, Gelpi E, Kouwenhoven M, Kros JM, Sanson M, Idbaih A, Brandes AA, Heinzl H, Gorlia T, Hainfellner JA, van den bent M. Prognostic value of Ki67 index in anaplastic oligodendroglial tumours--a translational study of the European Organization for Research and Treatment of Cancer brain tumor group. Histopathology. 2012;60(6):885–94.
16.
go back to reference Nielsen LAG, Bangsø JA, Lindahl KH, Dahlrot RH, Hansen S, Kristensen BW. Evaluation of the proliferation marker Ki-67 in gliomas: interobserver variability and digital quantification. Clin Neuropathol. 2015;34(3):157. Nielsen LAG, Bangsø JA, Lindahl KH, Dahlrot RH, Hansen S, Kristensen BW. Evaluation of the proliferation marker Ki-67 in gliomas: interobserver variability and digital quantification. Clin Neuropathol. 2015;34(3):157.
17.
go back to reference Music D, Dahlrot RH, Hermansen SK, Hjelmborg J, de Stricker K, Hansen S, Kristensen BW. Expression and prognostic value of the WEE1 kinase in gliomas. J Neuro-Oncol. 2016;127(2):381–9. Music D, Dahlrot RH, Hermansen SK, Hjelmborg J, de Stricker K, Hansen S, Kristensen BW. Expression and prognostic value of the WEE1 kinase in gliomas. J Neuro-Oncol. 2016;127(2):381–9.
18.
go back to reference Petterson SA, Dahlrot RH, Hermansen SK, KAM S, Gundesen MT, Wohlleben H, Rasmussen T, Beier CP, Hansen S, Kristensen BW. High levels of c-met is associated with poor prognosis in glioblastoma. J Neuro-Oncol. 2015;122(3):517–27. Petterson SA, Dahlrot RH, Hermansen SK, KAM S, Gundesen MT, Wohlleben H, Rasmussen T, Beier CP, Hansen S, Kristensen BW. High levels of c-met is associated with poor prognosis in glioblastoma. J Neuro-Oncol. 2015;122(3):517–27.
19.
go back to reference Dahlrot RH, Hansen S, Jensen SS, Schroder HD, Hjelmborg J, Kristensen BW. Clinical value of CD133 and nestin in patients with glioma: a population-based study. Int J Clin Exp Pathol. 2014;7(7):3739–51. Dahlrot RH, Hansen S, Jensen SS, Schroder HD, Hjelmborg J, Kristensen BW. Clinical value of CD133 and nestin in patients with glioma: a population-based study. Int J Clin Exp Pathol. 2014;7(7):3739–51.
20.
go back to reference Dahlrot RH, Hansen S, Herrstedt J, Schroder HD, Hjelmborg J, Kristensen BW. Prognostic value of Musashi-1 in gliomas. J Neuro-Oncol. 2013;115(3):453–61. Dahlrot RH, Hansen S, Herrstedt J, Schroder HD, Hjelmborg J, Kristensen BW. Prognostic value of Musashi-1 in gliomas. J Neuro-Oncol. 2013;115(3):453–61.
21.
go back to reference Dahlrot RH, Kristensen BW, Hjelmborg J, Herrstedt J, Hansen S. A Population-based study of high-grade gliomas and mutated isocitrate dehydrogenase 1. Int J Clin Exp Pathol. 2013;6(1):31–40. Dahlrot RH, Kristensen BW, Hjelmborg J, Herrstedt J, Hansen S. A Population-based study of high-grade gliomas and mutated isocitrate dehydrogenase 1. Int J Clin Exp Pathol. 2013;6(1):31–40.
22.
go back to reference Dahlrot RH, Kristensen BW, Hjelmborg J, Herrstedt J, Hansen S. A population-based study of low-grade gliomas and mutated isocitrate dehydrogenase 1 (IDH1). J Neuro-Oncol. 2013;114(3):309–17. Dahlrot RH, Kristensen BW, Hjelmborg J, Herrstedt J, Hansen S. A population-based study of low-grade gliomas and mutated isocitrate dehydrogenase 1 (IDH1). J Neuro-Oncol. 2013;114(3):309–17.
23.
go back to reference Kirkwood BR, Sterne JAC. Essential Medical Statistics. 2nd ed. Oxford: Blackwell Science; 2003. Kirkwood BR, Sterne JAC. Essential Medical Statistics. 2nd ed. Oxford: Blackwell Science; 2003.
24.
27.
go back to reference Prayson RA, Castilla EA, Hembury TA, Liu W, Noga CM, Prok AL. Interobserver variability in determining MIB-1 labeling indices in oligodendrogliomas. Ann Diagn Pathol. 2003;7(1):9–13. Prayson RA, Castilla EA, Hembury TA, Liu W, Noga CM, Prok AL. Interobserver variability in determining MIB-1 labeling indices in oligodendrogliomas. Ann Diagn Pathol. 2003;7(1):9–13.
28.
go back to reference Preusser M, Heinzl H, Gelpi E, Hoftberger R, Fischer I, Pipp I, Milenkovic I, Wohrer A, Popovici F, Wolfsberger S, Hainfellner JA. Ki67 index in intracranial ependymoma: a promising histopathological candidate biomarker. Histopathology. 2008;53(1):39–47. Preusser M, Heinzl H, Gelpi E, Hoftberger R, Fischer I, Pipp I, Milenkovic I, Wohrer A, Popovici F, Wolfsberger S, Hainfellner JA. Ki67 index in intracranial ependymoma: a promising histopathological candidate biomarker. Histopathology. 2008;53(1):39–47.
29.
go back to reference Coleman KE, Brat DJ, Cotsonis GA, Lawson D, Cohen C. Proliferation (MIB-1 expression) in oligodendrogliomas: assessment of quantitative methods and prognostic significance. Appl Immunohistochem Mol Morphol. 2006;14(1):109–14. Coleman KE, Brat DJ, Cotsonis GA, Lawson D, Cohen C. Proliferation (MIB-1 expression) in oligodendrogliomas: assessment of quantitative methods and prognostic significance. Appl Immunohistochem Mol Morphol. 2006;14(1):109–14.
30.
go back to reference Lopez XM, Debeir O, Maris C, Rorive S, Roland I, Saerens M, Salmon I, Decaestecker C. Clustering methods applied in the detection of Ki67 hot-spots in whole tumor slide images: an efficient way to characterize heterogeneous tissue-based biomarkers. Cytometry A. 2012;81(9):765–75. Lopez XM, Debeir O, Maris C, Rorive S, Roland I, Saerens M, Salmon I, Decaestecker C. Clustering methods applied in the detection of Ki67 hot-spots in whole tumor slide images: an efficient way to characterize heterogeneous tissue-based biomarkers. Cytometry A. 2012;81(9):765–75.
31.
go back to reference Swiderska Z, Korzynska A, Markiewicz T, Lorent M, Zak J, Wesolowska A, Roszkowiak L, Slodkowska J, Grala B. Comparison of the manual, semiautomatic, and automatic selection and leveling of hot spots in whole slide images for Ki-67 quantification in meningiomas. Anal Cell Pathol (Amst). 2015;2015:498746. Swiderska Z, Korzynska A, Markiewicz T, Lorent M, Zak J, Wesolowska A, Roszkowiak L, Slodkowska J, Grala B. Comparison of the manual, semiautomatic, and automatic selection and leveling of hot spots in whole slide images for Ki-67 quantification in meningiomas. Anal Cell Pathol (Amst). 2015;2015:498746.
Metadata
Title
Evaluation of the proliferation marker Ki-67 in gliomas: Interobserver variability and digital quantification
Authors
Ljudmilla A. G. Nielsen
Julie A. Bangsø
Kim H. Lindahl
Rikke H. Dahlrot
Jacob v. B. Hjelmborg
Steinbjørn Hansen
Bjarne W. Kristensen
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Diagnostic Pathology / Issue 1/2018
Electronic ISSN: 1746-1596
DOI
https://doi.org/10.1186/s13000-018-0711-2

Other articles of this Issue 1/2018

Diagnostic Pathology 1/2018 Go to the issue