Swelling and replicative DNA synthesis of detergent-treated mouse ascites sarcoma cells

Abstract

Previous investigation showed that mouse ascites sarcoma cells permeabilized with appropriate concentrations of detergents (Triton X-100, Nonidet P-40 and Brij 58) had high replicative DNA synthesis in the presence of the four deoxyribonucleoside triphosphates, ATP, Mg2+ and proper ionic environment. The present study showed the optimum detergent concentration for DNA synthesis coincided closely with the minimum detergent concentration for inducing cell swelling. Phase contrast microscopy and electron microscopy of Triton-permeabilized cells showed the characteristic swollen cytoplasms and nucleus. Autoradiographic study showed that the DNA synthesis in permeable cells was confined to the nucleus. Cell viability and [3H] deoxythymidine uptake were impaired at much lower concentrations of Triton X-100 than the optimum concentration for in vitro DNA synthesis. In Triton-permeabilized cells, the minimum Triton concentration that produced cell swelling also seemed to produce high repliative DNA synthesis, which reflects the in vivo state of DNA synthesis.