Stimulation of cis-diamminedichloroplatinum(II) accumulation by modulation of passive permeability with genistein: an altered response in accumulation-defective resistant cells.

The effect of the tyrosine kinase inhibitor genistein on the accumulation of cisplatin (DDP) was investigated in DDP-sensitive and -resistant human 2008 ovarian carcinoma cell lines. DDP accumulation after a 1-h exposure was maximally increased by concurrent 40 micrometer genistein. The maximal stimulation of accumulation was observed after 2 h of total genistein exposure and was 83 +/- 13% (n = 5) higher than controls. With resistant C13() cells, however, the stimulation of accumulation was delayed until 4 h and was increased only 46 +/- 18% compared to controls. Revertant RH4 cells that retained the accumulation defect behaved like the C13() cells. Genistein stimulated [3H]mannitol accumulation (a marker of passive permeability) by 43 +/- 9% (n = 3) in 2008 cells, and the effect was maximal after 2 h of total genistein exposure. Changes in [3H]mannitol accumulation in 2008 parent cells were highly correlated with DDP accumulation (r = 0.9010). These experiments also revealed that [3H]mannitol accumulation after 2 h in C13() cells was reduced 38% compared to 2008 cells, a decrease that reflected the DDP accumulation defect. Fluid-phase pinocytosis determined with lucifer yellow CH as a marker showed no difference between 2008 and C13() cells and no effect of genistein. Genistein was demonstrated to clearly inhibit protein-tyrosine phosphorylation initiated by the epidermal growth factor receptor kinase. Differences were noted in the phosphotyrosine pattern between the 2008 and C13() cells. Under the conditions that had the maximal effect on DDP accumulation in 2008 cells, genistein decreased the IC50 of DDP 8.2-fold in 2008 cells and 4.7-fold in C13() cells. We conclude that: (a) genistein stimulates DDP accumulation by modulating the passive permeability of the plasma membrane; (b) C13() cells are less permeable to passively diffusing small molecules, which offers a mechanism for the DDP accumulation defect without invoking carrier proteins; (c) the effect of tyrosine kinase inhibition on passive permeability is altered in C13() cells; and (d) pinocytosis contributes insignificantly to DDP accumulation. Genistein, a dietary isoflavone, thus seems to be a promising clinical candidate for combination with DDP.