Inhibition of unscheduled DNA synthesis in human lymphocytes by chemical carcinogens

Abstract

Freshly isolated human peripheral lymphocytes were treated with an alkylating agent immediately after collection and subsequently treated with UV radiation. This system was used because it represents a method for assaying damage in cells immediately after their removal from the host. The amount of UV-induced repair was measured as unscheduled DNA synthesis (UDS) by incorporation of [3H]deoxythymidine into the cellular DNA. The alkylating agents beta-propiolactone (BPL) and methyl methane-sulfonate (MMS) inhibited UDS at concentrations of 0.08 mM and 0.6 mM, respectively. Lower concentrations had no effect. Lymphocytes allowed to remain in culture medium after treatment with the alkylating agents did not recover the ability to perform UV-induced UDS even when cells were irradiated 48 h after carcinogen treatment. The decrease in UV-induced UDS resulting from alkylating agent treatment could not be attributed to cell death.