Potentiation of Azidothymidine Cytotoxicity in Cisplatin-resistant Human Ovarian Carcinoma Cells

Abstract

The combination of cisplatin and AZT was synergistic in a subline (A2780DDP) of human ovarian carcinoma cells resistant to cisplatin, in contrast to the parental carcinoma cell line, A2780S. A2780DDP cells have elevated levels of enzymes necessary for DNA synthesis and repair. If A2780DDP cells respond to cisplatin with an increase in thymidine kinase activity, then AZT may chain terminate newly synthesized DNA. To test this hypothesis, a dual label [( 14C]-thymidine/[3H]-AZT) experiment was designed. A2780 cells were first incubated with [14C]-thymidine to label DNA and measure DNA degradation in response to cisplatin. These [14C]-thymidine labeled A2780 cells were then incubated for short intervals with [3H]-AZT to measure chain termination after cisplatin addition. A2780S cells responded to cisplatin with a modest increase in thymidine turnover and an increase in AZT incorporation. In contrast, A2780DDP cells initially responded to cisplatin treatment with a significant increase in thymidine turnover and a corresponding increase in [3H]-AZT incorporation. This was concomitantly associated with an increase in thymidine kinase mRNA within 2 hr after cisplatin treatment. Thus, the A2780DDP cells had the ability to rapidly turnover DNA, a property effectively exploited by the deoxythymidine analogue, AZT, utilizing the enhanced enzymes of the DNA synthesis and repair complex in A2780DDP cells.