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A novel quantitative PCR of proliferation markers (Ki-67, topoisomerase IIα, and TPX2): an immunohistochemical correlation, testing, and optimizing for mantle cell lymphoma

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Abstract

A clinical course of patients with mantle cell lymphoma (MCL) is aggressive, and the disease is rarely curable. Proliferation rate is the most important prognostic factor. We developed a novel, reliable, rapid, and routinely applicable approach allowing a precise quantitative assessment of three proliferation markers, Ki-67, topoisomerase IIα, and TPX2. A total of 95 lymphoma specimens were measured in the study by real-time reverse transcription PCR (RQ-RT-PCR). We tested the reproducibility and accuracy of the assay and correlated the results with the immunohistochemical staining of the corresponding proteins. The results obtained indicated individual variability of the mRNA expression levels, reflecting heterogeneity of the proliferation rate in individual patients. In general, we observed the highest mRNA expression in the group of Burkitt lymphomas and the lowest in patients with reactive lymphadenopathies. We found increased proliferation rate in MCLs with high cyclin D1 mRNA, indicating a quantitative control of the cell cycle. We observed a correlation between mRNA expression level and the immunohistochemical staining of corresponding proteins, which significantly argues for the prognostic significance of the mRNA expression measuring. We confirmed the accuracy of the current assay for a precise quantitative examination of the proliferation activity. Real-time RT-PCR provides a novel approach applicable for clinical trials, and it represents a potent approach allowing to stratify MCL patients for entry into clinical trials according to the expression of the proliferation signature genes in their tumors. This approach may contribute to improved and individualized therapeutic options respecting the individual progression risk of patients with MCL.

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Acknowledgments

This work was supported by the Internal grant of faculty hospital in Motol no. 9756, Grant GAUK 46/2006/C/2.LF, and the Research project of the Ministry of Health no. 00064203/6704. We thank Dr. W. Klapper and the late Prof. Dr. R. Parwaresh (Department of Hematopathology and Lymph Node Registry, University Hospitals of Schleswig-Holstein, Kiel, Germany) who kindly provided us a sample of the antibody Ki-S2 against TPX2.

Conflict of interests

The authors declare that they have no conflict of interest in this study.

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Authors and Affiliations

  1. Department of Pathology and Molecular Medicine, 2nd Medical School, Charles University in Prague and Faculty Hospital in Motol, V Úvalu 84, 150 06, Prague 5, Czech Republic

    Helena Brizova, Marketa Kalinova, Lenka Krskova, Marcela Mrhalova & Roman Kodet

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  1. Helena Brizova
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  2. Marketa Kalinova
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  3. Lenka Krskova
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  4. Marcela Mrhalova
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  5. Roman Kodet
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Correspondence to Helena Brizova.

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Brizova, H., Kalinova, M., Krskova, L. et al. A novel quantitative PCR of proliferation markers (Ki-67, topoisomerase IIα, and TPX2): an immunohistochemical correlation, testing, and optimizing for mantle cell lymphoma. Virchows Arch 456, 671–679 (2010). https://doi.org/10.1007/s00428-010-0922-8

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