Open Access Research

Cancer-associated fibroblasts from human NSCLC survive ablative doses of radiation but their invasive capacity is reduced

Turid Hellevik1, Ingvild Pettersen2, Vivian Berg2, Jan Olof Winberg3, Bjørn T Moe5, Kristian Bartnes4, Ruth H Paulssen2, Lill-Tove Busund35, Roy Bremnes12, Anthony Chalmers6 and Iñigo Martinez-Zubiaurre2*

Author Affiliations

1 Department of Oncology, University Hospital of Northern Norway, 9038 Tromsø, Norway

2 Department of Clinical Medicine, University of Tromsø, 9037, Tromsø, Norway

3 Department of Medical Biology, University of Tromsø, 9037 Tromsø, Norway

4 Department of Cardiothoracic and Vascular Surgery, University Hospital of Northern Norway, 9038 Tromsø, Norway

5 Department of Clinical Pathology, University Hospital of Northern Norway, 9038 Tromsø, Norway

6 Institute of Cancer Sciences, University of Glasgow, Glasgow, UK

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Radiation Oncology 2012, 7:59  doi:10.1186/1748-717X-7-59

Published: 13 April 2012

Abstract

Background

Cancer-Associated Fibroblasts (CAFs) are significant components of solid malignancies and play central roles in cancer sustainability, invasion and metastasis. In this study we have investigated the invasive capacity and matrix remodelling properties of human lung CAFs after exposure to ablative doses of ionizing radiation (AIR), equivalent to single fractions delivered by stereotactic ablative radiotherapy (SART) for medically inoperable stage-I/II non-small-cell lung cancers.

Methods

CAFs were isolated from lung tumour specimens from 16 donors. Initially, intrinsic radiosensitivity was evaluated by checking viability and extent of DNA-damage response (DDR) at different radiation doses. The migrative and invasive capacities of CAFs were thereafter determined after a sub-lethal single radiation dose of 18 Gy. To ascertain the mechanisms behind the altered invasive capacity of cells, expression of matrix metalloproteinases (MMPs) and their endogenous inhibitors (TIMPs) were measured in the conditioned media several days post-irradiation, along with expression of cell surface integrins and dynamics of focal contacts by vinculin-staining.

Results

Exposing CAFs to 1 × 18 Gy resulted in a potent induction of multiple nuclear DDR foci (> 9/cell) with little resolution after 120 h, induced premature cellular senescence and inhibition of the proliferative, migrative and invasive capacity. AIR promoted MMP-3 and inhibited MMP-1 appearance to some extent, but did not affect expression of other major MMPs. Furthermore, surface expression of integrins α2, β1 and α5 was consistently enhanced, and a dramatic augmentation and redistribution of focal contacts was observed.

Conclusions

Our data indicate that ablative doses of radiation exert advantageous inhibitory effects on the proliferative, migratory and invasive capacity of lung CAFs. The reduced motility of irradiated CAFs might be a consequence of stabilized focal contacts via integrins.

Keywords:
Cancer-associated fibroblasts; Ablative radiation; Invasion; Integrins; Focal adhesion