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Cells were irradiated using the clinical protocol RTS and recovered for comparison after rinsing in drug-free medium for 1 hour, then harvested for analysis at various time points. In the recovery experiments, however, no wash step was performed.
There was a pronounced effect of deoxycholate on cell survival. Comparison of cell survival curves for RTS and wash protocol revealed a similar magnitude of repopulation post-irradiation in untreated cells. This effect was dampened in cells exposed to RTS or a wash procedure, though SCC irradiated and washed with drug-free medium consistently displayed greater initial survival.
Repopulation on RTS and a wash protocol is increased with deoxycholate, which is similarly represented by demonstrating the increase survival of cells treated with the wash protocol relative to untreated controls. This, in addition to deoxycholate exposure inducing clustered cell death, indicates that deoxycholate plays a key role in promoting SCC repopulation.
Beyond demonstrating a measureable effect of drug on cell survival, the authors fitted the data to determine the relative repopulation rates by fitting a curve to the curves. Their analysis concluded that recovery consistently occurred at a lower rate in cells exposed to RTS or a wash protocol in the short-term [survival] experiments. For long-term [survival] experiments, no decrease in repopulation was found in cells exposed to a wash protocol, though the magnitude of the decrease in repopulation in cells exposed to RTS was similar.
The authors therefore conclude that recovery of tumour cells post-irradiation will be most effective when the first radiation session is executed in the absence of deoxycholate, though this will require considerable experimental variation and subsequent adjustments. The relationships observed between deoxycholate exposure and repopulation, as well as those between drug exposure and repopulation, suggest that concerns about the negative effect of drug on the reduction of repopulation following RTS may be unfounded. d2c66b5586
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