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The effect of cilengitide in combination with irradiation and chemotherapy in head and neck squamous cell carcinoma cell lines

Die Wirkung von Cilengitide in Kombination mit Bestrahlung und Chemotherapie bei Zelllinien des Kopf-Hals-Plattenepithelkarzinoms

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Abstract

Background

Integrins are highly attractive targets in oncology due to their involvement in angiogenesis in a wide spectrum of cancer entities. Among several integrin inhibitors under clinical evaluation, cilengitide is the most promising compound. However, little is known about the cellular processes induced during cilengitide therapy in combination with irradiation and cisplatin in head and neck squamous cell carcinoma (HNSCC).

Materials and methods

The cytostatic effect of cilengitide was assessed by proliferation assay in the three HNSCC cell lines SCC25, FaDu and CAL27. Combination experiments with cisplatin and irradiation were performed. Possible synergistic effects were calculated in combination index (CI) analyses. Colony forming inhibition was investigated in clonogenic assays. Real-time PCR arrays were used to evaluate target protein gene expression patterns. Flow cytometry was used to detect apoptosis.

Results

Used alone, cilengitide has only minor cytotoxic effects in HNSCC cell lines. However, combination with cisplatin resulted in synergistic growth inhibition in all three cell lines. Irradiation showed synergism in short-term experiments and in colony forming assays, an additive effect was detected. Real-time PCR assay detected downregulation of the antiapoptotic protein Bcl-2 after exposure of cells to cilengitide.

Conclusion

Cilengitide in combination with cisplatin and irradiation may be a feasible option for the treatment of patients with head and neck cancer. However, further investigations are required to understand the exact mechanism that leads to synergistic cytotoxicity.

Zusammenfassung

Hintergrund

Durch ihre Rolle bei der Angiogenese sind Integrine ein attraktives Ziel in der onkologischen Forschung. Der derzeit vielversprechendste Inhibitor dieser Moleküle ist Cilengitide, welches bereits in klinischen Studien getestet wird. Dennoch ist erst wenig über die zellulären Vorgänge bekannt, welche durch Cilengitide in Kopf-Hals-Karzinomen (HNSCC) insbesondere in Kombination mit Strahlentherapie und Cisplatin ausgelöst werden.

Material und Methoden

Der zytostatische Effekt von Cilengitide wurde in drei Kopf-Hals-Zelllinien SCC25, CAL27 und FaDu mittels Proliferationsassay überprüft. Mit Cisplatin und Bestrahlung wurden Kombinationsexperimente durchgeführt. Mögliche Synergien wurden mittels Kombinationsindex-(CI-)Analyse berechnet. Der Einfluss auf die Koloniebildung wurde in Koloniebildungsexperimenten gezeigt. In Real-time-PCR-Arrays wurden die Expressionsmuster von bekannten Zielproteinen untersucht. Mittels FACS („flow cytometry“) wurde nach Apoptose gesucht.

Resultate

Cilengitide selbst hatte nur eine geringe direkte Wirkung auf das Wachstum unserer Zelllinien. Die Kombination mit Cisplatin jedoch zeigte eine synergistische Wachstumshemmung. In Kurzzeitexperimenten war Cilengitide in Kombination mit Bestrahlung synergistisch. Auch in den Koloniebildungsassays konnte ein additiver Effekt nachgewiesen werden. Das antiapoptotische Bcl-2-Protein wurde nach Exposition mit Cilengitide vermindert exprimiert.

Schlussfolgerung

Cilengitide in Kombination mit Cisplatin und Bestrahlung könnte eine sinnvolle Behandlungsoption für Patienten mit Kopf-Hals-Karzinomen sein. Dafür sind jedoch noch weitere Experimente notwendig, auch um den exakten Mechanismus der Synergie zu verstehen.

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Conflict of interest

G. Heiduschka, C. Lill, S. Schneider, R. Seemann, G. Kornek, R. Schmid, U. Kotowski and D. Thurnher state that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Otorhinolaryngology, Head and Neck Surgery, Comprehensive Cancer Center, Medical University of Vienna, Waehringer Guertel 18–20, 1090, Vienna, Austria

    G. Heiduschka, C. Lill, S. Schneider, U. Kotowski & D. Thurnher M. D.

  2. Clinical Pharmacology, Medical University of Vienna, Vienna, Austria

    G. Heiduschka

  3. Craniomaxillofacial and Oral Surgery, Medical University of Vienna, Vienna, Austria

    R. Seemann

  4. Internal Medicine, Medical University of Vienna, Vienna, Austria

    G. Kornek

  5. Radiotherapy and Radiobiology, Medical University of Vienna, Vienna, Austria

    R. Schmid

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  1. G. Heiduschka
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Correspondence to D. Thurnher M. D..

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Heiduschka, G., Lill, C., Schneider, S. et al. The effect of cilengitide in combination with irradiation and chemotherapy in head and neck squamous cell carcinoma cell lines. Strahlenther Onkol 190, 472–479 (2014). https://doi.org/10.1007/s00066-014-0600-x

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  • DOI: https://doi.org/10.1007/s00066-014-0600-x

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