IntroductionThe success of radiotherapy depends on the ability to inhibit tumour growth, and relapse after therapy is determined by cells that retain their clonogenic potential. The radiation sensitivity of isolated tumour cell clones in vitro is routinely determined with clonogenic assays. In solid tumours, however, clonogenic cells are not isolated and we carried out experiments to measure the influences of cell-cell contact on their proliferative potential. To this end we developed a new experimental approach to measure the effects of radiation on tumour cell populations. The observations can be understood with the help of a novel stochastic model with a well-defined biological basis.Material and methodsT47D cells (human breast carcinoma) were grown at various concentrations in F(flat)-bottom and V-bottom wells of 96-well culture plates. The spheroid outgrowth method was also used to obtain densely-packed tissue cell cultures. A Gammacell40 irradiator equipped with a 137Cs source was used to treat cell cultures. Cell fusion was assessed by confocal microscopy. Syncytin 1 expression was assessed by RT-PCR and by flow cytometry using an anti-HERV antibody (clone ab7115, Abcam).Results and discussionsThe probability of cell survival after 8 Gy radiation treatment increased ~4.7 times when the cells were grown in V-bottom wells as compared to cells grown in F-bottom wells (p(survival)=0.0113 and 0.0024, respectively). Microscopic inspections of tissue-like cultures showed that after treatment cell populations were mostly composed of giant cells with multiple nuclei. Cytoplasmic bridges joining different cells were clearly visible. Giant cells and cytoplasmic bridges disappeared at later times (>600 hours) when the cells displayed normal morphology and started to proliferate again. Sequence analysis of cloned RT-PCR products showed that cells expressed a Syncytin 1 homologous protein (Sp). Flow cytometry assays confirmed cytoplasmic expression of Sp and revealed that Sp translocated to the cell surface of irradiated cells committed to death. The fraction of cells surviving 8 Gy treatment was significantly reduced in cultures treated with anti-Sp antibodies.ConclusionOur experimental findings indicate that recovery of breast tumours from radiation is very likely to involve complex pathways that act at the cell population level and that include events of cell fusion mediated by a protein homologous to Syncytin 1.