Response of Human Squamous Cell Carcinoma Xenografts of Different Sizes to Irradiation: Relationship of Clonogenic Cells, Cellular Radiation Sensitivity in Vivo, and Tumor Rescuing Units

The relationship of clonogenic cells, cellular radiation sensitivity at tumor control doses in vivo, and tumor rescuing units at different tumor sizes was investigated in the human squamous cell carcinoma FaDu growing in NCr/Sed nude mice. The composition of the tumors was determined in single cell suspensions and compared to tumor control data after single-dose irradiation. To avoid the influence of varying oxygen concentrations in the tumors, all irradiations were performed under clamp hypoxia. Nude mice and animals further immunosuppressed by 6-Gy whole-body irradiation were used to assess the immunological effects. The numbers of total cells, cells excluding trypan blue, host cells, and colony-forming cells increased linearly with the weight of FaDu tumors. Comparable results were obtained for cell suspensions prepared from tumors growing in nude or pretreated nude mice. The radiation dose required to control 50% of tumors (<tex-math>${\rm TCD}_{50}$</tex-math>) of different sizes between 36 and <tex-math>$470\ {\rm mm}^{3}$</tex-math> increased from 52.1 to 60.1 Gy when the tumors were maintained in normal nude mice and from 50.8 to 61.3 Gy in whole-body-irradiated mice. The D₀ of FaDu cells in vivo was calculated by regression analysis of <tex-math>${\rm TCD}_{50}$</tex-math> vs the logarithm of the clonogenic cell number, assuming an oxygen enhancement ratio of 3.0. The resultant <tex-math>$D_{0}{\rm s}$</tex-math> of 1.1 and 1.2 Gy in vivo correspond well to the radiosensitivity of FaDu cells in vitro determined previously. Assuming the single-hit multitarget model of cell killing and extrapolation numbers between 2 and 20, the mean number of tumor rescuing units would be 10⁵ to 10⁶ for a <tex-math>$100\text{-}{\rm mm}^{3}$</tex-math> tumor growing in whole-body-irradiated nude mice. Comparison of the number of tumor rescuing units to the estimated number of clonogenic cells does not conflict with the assumption that every surviving clonogenic cell is able to repopulate FaDu tumors after irradiation; however, it seems more likely that more than one clonogenic cell is necessary. The proportion of tumor rescuing units in the clonogenic cell population is independent of tumor size.