Effects of adriamycin and hyperthermia on cellular uptake of [³H]thymidine and its significance for the incorporation into DNA

Abstract

Inhibition of the salvage DNA synthesis was studied in ascites tumour cells up to 240 min at moderately increased temperatures of 39°C, 40.5°C and 43°C alone or in combination with adriamycin (ADR). Hyperthermia and ADR acted additively on salvage DNA synthesis. The [³H]thymidine (TdR) incorporation was strongly related to the cellular uptake of [³H]TdR. In order to evaluate the significance of the decrease in [³H]TdR uptake for the incorporation into DNA, the thymidine kinase (TK) has also been studied. TK activity started to decrease at a temperature of 39°C, probably due to inhibition of translation and/or transcription of the enzyme, and was almost completely inhibited at 43°C. ADR did not affect TK activity. Inhibition of DNA synthesis by aphidicolin neither decreased the ability of the cell to take up [³H]TdR nor the ability to affect TK activity. We concluded that moderate increases in temperature alone, or in combination with ADR, inhibit salvage DNA synthesis by inhibition of TdR uptake, possibly due to inhibition of phosphorylation; ADR, on the contrary, inhibits TdR uptake by other mechanisms such as changes in membrane function.

Key Words:

• adriamycin
• hyperthermia
• thymidine uptake and incorporation
• thymidine kinase.