Transfer of growth inhibition between normal and virus-transformed cells: autoradiographic studies using marked cells

Abstract

[³H]Thymidine and [³H]hypoxanthine incorporation were investigated by autoradiography in mixed cultures of polyoma-transformed BHK21 cells and freshly isolated mouse fibroblasts, with ingested carbon or carmine granules as markers to distinguish the cells. An assessment of the marking technique showed that there was some exchange of granules in the mixed cultures which prevented certain identification of individual cells, but suitable criteria were chosen for distinguishing the cells on a statistical basis. Thymidine incorporation was inhibited in one third to two thirds of the transformed cells when they were in contact with stationary layers of normal cells, which themselves showed a low proportion with thymidine incorporation. Transformed cells in the same dish which were not touching the normal cells showed no inhibition of thymidine incorporation. This is in agreement with the earlier observation that growth of transformed BHK21 cells is inhibited by contact with stationary normal fibroblasts. Experiments were also carried out on hypoxanthine incorporation with the TG1 variant of polyoma-transformed cells. TG1 cells are deficient in inosinic pyrophosphorylase and autoradiography shows a failure to incorporate hypoxanthine. When TG1 cells were cultured in contact with normal mouse embryo cells, however, it was found that hypoxanthine was present in the TG1 cells as well as in the normal cells. Increased incorporation did not occur in TG1 cells in the same dish which were not in contact with normal cells. This confirms earlier observations and shows that certain substances can pass directly from normal to transformed cells. It suggests the possibility that molecules concerned in growth regulation might also be transferred directly between contiguous cells.