NOVOBIOCIN-INDUCED ACCUMULATION OF ETOPOSIDE (VP-16) IN WEHI-3B D+ LEUKEMIA-CELLS

作者：LORICO A; RAPPA G; SARTORELLI AC^*

来源：International Journal of Cancer, 1992, 52(6): 903-909.

DOI：10.1002/ijc.2910520613

A previous report from this laboratory demonstrated that novobiocin produced supra-additive cytotoxicity when combined with etoposide(VP-16)orteniposide(VM-26) in WEHI-3B D+ and A549 cells. The increase in cytotoxicity was accompanied by an increase in the formation of drug-stabilized protein-DNA covalent complexes. We now report that novobiocin increased the amount of VP-16-induced covalent complexes between the 170 kDa form of topoisomerase II and DNA in WEHI-3B D+ cells, as measured by the band-depletion immunoblotting assay, while it did not affect the extractable topoisomerase II activity, measured by the unknotting of P4 phage DNA and by a DNA cleavage assay. Novobiocin progressively increased the steady-state concentration of intracellular VP-16. Removal of novobiocin resulted in a rapid return of VP-16 to levels comparable to those seen with VP-16 alone. The increased accumulation of VP-16 was accounted for by an increase in the exchangeable fraction only. The novobiocin-mediated increase in the steady-state concentration of VP-16 occurred whether novobiocin was added simultaneously with VP-16 or was added after a steady-state level of VP-16 had been achieved. Novobiocin did not affect the initial rate of uptake of VP-16; however, it inhibited the efflux of the epipodophyllotoxin. In fact, when cells were loaded with the same level of VP-16 in the presence or absence of novobiocin, the efflux curves in the presence or absence of novobiocin were significantly different. We conclude that the inhibition of VP-16 efflux by novobiocin is responsible for the increase in VP-16 accumulation, leading to increased formation of VP-16-stabilized topoisomerase-II-DNA covalent complexes and increased cytotoxicity.

• 出版日期1992-12-2