3D cell cultures of human head and neck squamous cell carcinoma cells are radiosensitized by the focal adhesion kinase inhibitor TAE226

doi:10.1016/j.radonc.2009.08.001

Radiotherapy and Oncology

Volume 92, Issue 3, September 2009, Pages 371-378

https://doi.org/10.1016/j.radonc.2009.08.001 Get rights and content

Abstract

Background and purpose

Focal adhesion kinase (FAK), a main player in integrin signaling and survival, is frequently overexpressed in human cancers and therefore postulated as potential target in cancer therapy. The aim of this study was to evaluate the radiosensitizing potential of the FAK inhibitor TAE226 in three-dimensional (3D) tumor cell cultures.

Materials and methods

Head and neck squamous cell carcinoma (HNSCC) cells (FaDu, UT-SCC15, UT-SCC45), lung cancer cells (A549), colorectal carcinoma cells (DLD-1, HCT-116) and pancreatic tumor cells (MiaPaCa2, Panc1) were treated with different concentrations of TAE226 (0–1 μm; 1 or 24 h) without or in combination with irradiation (0–6 Gy, X-ray, single dose). Subsequently, 3D clonogenic survival assays (laminin-rich extracellular matrix) and Western blotting (expression/phosphorylation, e.g. FAK, Akt, ERK1/2) were performed.

Results

All investigated 3D cell cultures showed a dose-dependent reduction in clonogenic survival by TAE226. Intriguingly, TAE226 only significantly radiosensitized 3D HNSCC cell cultures accompanied by a pronounced dephosphorylation of FAK, Akt and ERK1/2.

Conclusions

Our data demonstrate TAE226 as potent FAK inhibitor that enhances the cellular radiosensitivity particularly of HNSCC cells grown in a 3D cell culture model. Future in vitro and in vivo investigations will clarify, to which extent this approach might be clinically relevant for radiotherapy of HNSCC.

Section snippets

Cell culture

UT-SCC15, UT-SCC45 (Ref. in [41]) and FaDu human head and neck squamous cell carcinoma (HNSCC) cell lines as well as A549 human lung cancer, DLD-1 and HCT-116 human colorectal carcinoma, MiaPaCa2 and Panc1 human pancreatic carcinoma cell lines were obtained from ATCC (Rockville, USA). Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; PAA, Cölbe, Germany) containing glutamax-I (l-alanyl-l-glutamine) supplemented with 10% fetal calf serum (FCS; PAA) and 1% non-essential amino acids

TAE226 reduces clonogenic survival dose- and time-dependently

Clonogenic survival was determined in 3D tumor cell cultures upon treatment with the FAK inhibitor TAE226 (Fig. 1a and b). Increasing concentrations of TAE226 significantly (p < 0.05) diminished clonogenic survival of 3D grown FaDu, UT-SCC15, UT-SCC45 (all HNSCC), A549 (lung cancer), DLD-1, HCT-116 (colorectal carcinoma) and MiaPaCa2 (pancreatic carcinoma) cells after a 1-h treatment in comparison to DMSO control (Fig. 1c). A 24-h treatment resulted in further reduction of clonogenicity in all

Discussion

The development and progression of SCC is thought to be critically dependent on integrins and their downstream signal transduction [46]. As FAK is well known to channel prosurvival cues from both integrins and growth factor receptors, we assessed, for the first time, the effects of pharmacological FAK inhibition using TAE226 in HNSCC in combination with irradiation, which is part of the standard therapy of this disease. The major findings generated in a more physiological 3D cell culture model

Acknowledgements

The research and the authors were in part supported by a Grant from the Bundesministerium für Bildung und Forschung (BMBF Contract 03ZIK041). The pharmacological FAK inhibitor TAE226 was kindly provided by Novartis (Switzerland).

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Molecular radiobiology3D cell cultures of human head and neck squamous cell carcinoma cells are radiosensitized by the focal adhesion kinase inhibitor TAE226

Abstract

Background and purpose

Materials and methods

Results

Conclusions

Section snippets

Cell culture

TAE226 reduces clonogenic survival dose- and time-dependently

Discussion

Acknowledgements

Radiother Oncol

Biochim Biophys Acta

Clin Oncol [R Coll Radiol]

Radiother Oncol

Exp Cell Res

Am J Pathol

J Biol Chem

J Biol Chem

J Biol Chem

J Biol Chem

J Biol Chem

Radiother Oncol

Radiother Oncol

Eur J Cancer Clin Oncol

Exploring the role of cancer stem cells in radioresistance

Nat Rev Cancer

Cancer stem cells: redefining the paradigm of cancer treatment strategies

Mol Interv

Tumour stem cells and drug resistance

Nat Rev Cancer

Targeting molecular determinants of tumor chemo-radioresistance

Semin Oncol

Cell adhesion-mediated radioresistance (CAM-RR) extracellular matrix-dependent improvement of cell survival in human tumor and normal cells in vitro

Strahlenther Onkol

Pharmacological inhibition of EGFR tyrosine kinase affects ILK-mediated cellular radiosensitization in vitro

Int J Radiat Biol

Focal adhesion kinase: targeting adhesion signaling pathways for therapeutic intervention

Clin Cancer Res

Immunohistochemical analyses of focal adhesion kinase expression in benign and malignant human breast and colon tissues: correlation with preinvasive and invasive phenotypes

Clin Cancer Res

Adhesion systems in normal breast and in invasive breast carcinoma

Am J Pathol

Overexpression of focal adhesion kinase (p125FAK) in human colorectal carcinoma liver metastases: independence from c-src or c-yes activation

Ann Surg Oncol

Molecular radiobiology
3D cell cultures of human head and neck squamous cell carcinoma cells are radiosensitized by the focal adhesion kinase inhibitor TAE226