Effect of serum and growth factors on heat sensitivity in Swiss mouse 3T3 cells

Quiescent Swiss mouse 3T3 cells react to a heat treatment at 46 degrees C for 20 min by changing their flat, well-extended morphology to a round appearance with retracted cytoplasmic processes during the subsequent 2 h at 37 degrees C. The percentage of morphologically changed cells was used to quantify changes in heat sensitivity, or resistance, in response to mitogenic stimulation. Stimulating quiescent cells with serum or with the specific growth factors epidermal growth factor (EGF) and prostaglandin F2 alpha (PGF2 alpha) markedly increased the heat resistance to a 46 degrees C treatment, but only when the heat treatment, but only when the heat treatment was applied within 2-3 h after the addition. When insulin (which is not mitogenic, but synergistic with EGF and PGF2 alpha in these cells) was added alone or in combination with either EGF or PGF2 alpha, it has no effect on the development of heat resistance. Neither did cycloheximide nor tunicamycin inhibit heat resistance induced by EGF, and cycloheximide even enhanced it after 2-4 h. However, adding colcemid before or at the beginning of the heat treatment abolished the increased heat resistance. The results indicate that the resistance to a single heat treatment at 46 degrees C may be related to changes in the metabolic state after mitogenic stimulation, even though these changes need not be reflected in the rate of entry into S phase. Furthermore, the cytoskeletal organization appears to be a crucial component in heat resistance of Swiss 3T3 cells.