Modulation of cisplatin cytotoxicity by permeabilization of the plasma membrane by digitonin in vitro.
Killing of human ovarian carcinoma 2008 cells by cisplatin (DDP) is in direct proportion to the amount of drug entering the cell. DDP and its analogue [3H]dichloro(ethylenediamine)platinum[II] ([3H]-DEP) enter cells relatively slowly. We found that the uptake of [3H]DEP into 2008 cells could be increased by treating the cells briefly with the plasma membrane-selective detergent digitonin. A similar effect was observed in an 11-fold DDP-resistant subline of 2008 cells, designated 2008/C13*5.25. A measurable effect was produced by concentrations as low as 5 microM, and 40 microM digitonin increased [3H]DEP accumulation at 1 hr by 4.4 +/- 0.2- and 6.5 +/- 0.7-fold (means +/- SD) in 2008 and 2008/C13*5.25 cells, respectively. The effect was rapid, occurring within 1 min. Increased [3H]DEP uptake was accompanied by increased platination of DNA (8.5-fold in 2008 cells and 18.5-fold in 2008/C13*5.25 cells), and by enhanced killing of both the DDP-sensitive and -resistant cells that was shown to be synergistic by median effect analysis. The combination index at 50% cell kill was 0.64 +/- 0.14 (values < 1 indicate synergy). We conclude that a brief exposure to digitonin can increase [3H]DEP uptake in vitro, and can overcome the impaired [3H]DEP accumulation associated with acquired DDP resistance. DDP and digitonin interact synergistically to increase tumor cell kill in vitro.
Department of Medicine, University of California, San Diego, La Jolla 92093-0812.
A P
AP
Jekunen
D R
DR
Shalinsky
D K
DK
Hom
K D
KD
Albright
D
D
Heath
S B
SB
Howell
D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000633:toxicity; D016923:Cell Death / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002945:Cisplatin / Q000493:pharmacokinetics* / Q000633:toxicity*; D004273:DNA, Neoplasm / Q000187:drug effects / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004072:Digitonin / Q000494:pharmacology*; D004357:Drug Synergism; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D009944:Organoplatinum Compounds / Q000493:pharmacokinetics; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000378:metabolism; D012680:Sensitivity and Specificity; D014316:Tritium; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D003902:Detergents; D009944:Organoplatinum Compounds; D014316:Tritium; C002892:platinum ethylenediamine dichloride; D004072:Digitonin; D002945:Cisplatin