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Influence of buffy coat–derived putative endothelial progenitor cells on tumor growth and neovascularization in oral squamous cell carcinoma xenografts

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Abstract

Objectives

The aim of this murine in vivo study was to investigate whether buffy coat–derived putative endothelial progenitor cells (BCEPC) alter tumor growth and neovascularization in oral squamous cell carcinomas (OSCC).

Materials and methods

A murine xenograft model using the PCI-13 oral cancer cell line was deployed of which n = 24 animals received 2 × 106 BCEPC by transfusion whereas the control group (n = 24) received NaCl (0.9%) instead. Tumor size, volume, and capillary density were determined by sonography and measurement with a caliper. Immunohistochemical analysis was carried out with antibodies specific for Cytokeratins, Flt-4, Podoplanin, and Vimentin.

Results

In the experimental group, systemic application of BCEPC significantly increased tumor volume to 362.49% (p = 0.0012) and weight to 352.38% (p = 0.0018) as well as vascular densities to 162.15% (p = 0.0021) compared with control tumors. In addition, BCEPC-treated xenografts exhibited higher Cytokeratin expression levels by a factor of 1.47 (p = 0.0417), Podoplanin by a factor of 3.3 (p = 0.0020) and Vimentin by a factor of 2.5 (p = 0.0001), respectively.

Conclusions

Immunohistochemical investigations support the notion that BCEPC transfusion influences neovascularization and lymphatic vessel density, thereby possibly promoting tumor progression. Future studies, which will include gene expression analysis, should help to define the possible role of BCEPC during OSCC progression in more detail.

Clinical relevance

Endothelial progenitor cells (EPCs) could serve as a target structure for the treatment of OSCC and possibly other solid tumors.

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Acknowledgements

The authors thank Dr. J. Goldschmitt (Laboratory of the Clinic for Oral and Maxillofacial Surgery, University Medical Center Mainz, Germany) as well as R. Peldszus and G. Sadowski (Interdisciplinary Head & Neck Oncology Laboratory, Department of Otolaryngology, University Hospital Marburg, Germany) for their excellent technical support. We also thank PD Dr. M. Bette (Institute of Anatomy and Cell Biology, Philipps-Universität Marburg) for the excellent support during microscopic evaluation. Finally, we also thank Dr. A. Schmidt and members of the Institute of Pathology and Molecular Pathology (University Hospital Marburg) for their collegial support and the provision of reagents. This study is part of the thesis work of M.O.

Funding

The study was funded by the foundation for tumor research head and neck (Stiftung für Tumorforschung Kopf-Hals), Wiesbaden, Germany.

Author information

Authors and Affiliations

  1. Interdisciplinary Head & Neck Oncology Laboratory, Department for Oral and Maxillofacial Surgery, University Hospital Giessen and Marburg, Campus Marburg, Baldingerstrasse, D-35033, Marburg, Germany

    Marius Otto, Andreas Neff & Thomas Ziebart

  2. Clinic for Oral and Maxillofacial Surgery, University Medical Center Mainz, Mainz, Germany

    Sebastian Blatt

  3. Clinic for Oral and Maxillofacial Surgery, Federal Armed Forces Hospital Koblenz, Koblenz, Germany

    Andreas Pabst

  4. Interdisciplinary Head & Neck Oncology Laboratory, Department of Otolaryngology, Head & Neck Surgery, University Hospital Marburg, Marburg, Germany

    Robert Mandic

  5. Department of Mathematics and Computer Science, Research Group for Bioinformatics, Philipps-Universität Marburg, Marburg, Germany

    Johanna Schwarz

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  1. Marius Otto
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Correspondence to Thomas Ziebart.

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The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The local ethics committee (Landesärztekammer Rheinland-Pfalz) approved all of the experiments with human material in this article (Ethikvotum 837.387.11 (7929)). All applicable international, national, and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. A corresponding animal test application (animal experiment G 12-1-037) for the experiments was approved by the Landesuntersuchungsamt Rheinland-Pfalz in Koblenz and thus meets all §8 Paragraph 3 (2) of the German Animal Welfare Act required preconditions.

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Otto, M., Blatt, S., Pabst, A. et al. Influence of buffy coat–derived putative endothelial progenitor cells on tumor growth and neovascularization in oral squamous cell carcinoma xenografts. Clin Oral Invest 23, 3767–3775 (2019). https://doi.org/10.1007/s00784-019-02806-2

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