Summary
We examined the short-term accumulation of cisplatin (DDP) in sensitive 2008 human ovarian carcinoma cells and in a 2- to 3-fold DDP-resistant and accumulation-deficient variant. During the 1st min of exposure to 500 μM DDP, sensitive cells accumulated platinum at a rate of 187±63 pmol/mg protein per min, whereas resistant cells accumulated platinum at 123±85 pmol/mg protein per min, a rate that was 66% that of sensitive cells. From 2–10 min of exposure, sensitive and resistant cells accumulated the drug at rates of 51.4±21.5 and 34.0±9.70 pmol/mg protein per min, respectively. In resistant cells, this rate again represented 66% that of sensitive cells. For each cell line, the DDP accumulation was 3.6 times faster during the 1st min that it was over 2–10 min. Initial DDP accumulation was linear with drug concentration in each cell line. Efflux measurements were made over a 50-min period after a 10-min load in 500 μM DDP. The loss of platinum was biphasic in each cell line, with an initial, rapidly effluxing component being lost within 10 min in each cell line. The rate constant for loss of platinum from this rapidly effluxing pool, measured after a 10-min loading period in 500 μM DDP, was 0.67±0.09 s−1 in sensitive cells and 1.03±0.15 s−1 (a 53% increase) in resistant cells. Between 5 and 50 min of an accumulation time course in 500 μM DDP, the size of the rapidly effluxing platinum pool remained relatively constant in each cell line, with the major contribution to the increase in total platinum over time coming from growth of the slowly effluxing platinum pool. We conclude that diminished retention of platinum in the rapidly effluxing pool of resistant cells in a major determinant of decreased DDP accumulation in these cells.
Similar content being viewed by others
Abbreviations
- DDP:
-
cis-diamminedichloroplatinum(II)
- DEP:
-
cis-dichloro(ethylenediamine)platinum(II)
- RPMI:
-
Roswell Park Memorial Institute
- PBS:
-
phosphate-buffered saline, consisting of (per liter) 8 g NaCl, 0.2 g KCl, 1.15 g Na2HPO4, and 0.2 g KH2PO4
References
Andrews PA, Murphy MP, Howell SB (1985) Differential potentiation of alkylating and platinating agent toxicity in human ovarian carcinoma cells by glutathione depletion. Cancer Res 45: 6250–6253
Andrews PA, Velury S, Mann SC, Howell SB (1988)cis-Diamminedichloroplatinum(II) accumulation in sensitive and resistant human ovarian carcinoma cells. Cancer Res 48: 68–73
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254
Disaia PJ, Sinkovics JG, Rutledge FN, Smith JP (1972) Cell-mediated immunity to human malignant cells. Am J Obstet Gynecol 114: 979–989
Eichholtz-Wirth H, Hietel B (1986) The relationship between cisplatin sensitivity and drug uptake into mamalian cells in vitro. Br J Cancer 54: 239–243
Gale GR, Morris CR, Atkins LM, Smith AB (1973) Binding of an antitumor compound to cells as influenced by physical factors and pharmacologically active agents. Cancer Res 33: 813–818
Golenberg GJ, Vanstone CL, Israels LG, Ilse D, Bihler I (1970) Evidence for a transport carrier of nitrogen mustard in nitrogen mustard-sensitive and-resistant L5178Y lymphoblasts. Cancer Res 30: 2285–2291
Goldman DI, Lichtenstein NS, Oliverio VT (1968) Carrier-mediated transport of the folic acid analogue, methotrexate, in the L1210 leukemia cells. J Biol Chem 243: 5007–5017
Gross RB, Scanlon KJ (1986) Amino acid membrane transport properties of L1210 cells resistant to cisplatin. Chemioterapia 5: 37–43
Hecquet B, Leroy A, Lefebvre JL, Peyrat JP, Adenis L (1986) Uptake of platinum compounds in human tumors. In vitro study. Bull Cancer (Paris) 73: 535–541
Howell SB, Pfeifle CL, Wung WE, Olshen RA, Lucas WE, Yon JL, Green M (1982) Intraperitoneal cisplatin with systemic thiosulfate protection. Ann Intern Med 97: 845–851
Hromas RA, North JA, Burns PC (1987) Decreased cisplatin uptake by resistant L1210 leukemia cells. Cancer Lett 36: 197–201
Kraker AJ, Moore CW (1988) Accumulation ofcis-diamminedichloroplatinum(II) and platinum analogues by platinum-resistant murine leukemia cells in vitro. Cancer Res 48: 9–13
Richon VM, Schulte N, Eastman A (1987) Multiple mechanisms of resistance tocis-diamminedichloroplatinum(II) in murine leukemia L1210 cells. Cancer Res 47: 2056–2061
Riordan JR, Ling V (1985) Genetic and biochemical characterization of multidrug resistance. Pharmacol Ther 28: 51–75
Schafer JA, Barfuss DW (1986) Mechanisms of transmembrane transport in isolated cells and their experimental study. In: Membrane transport of antineoplastic agents. Pergamon, New York, pp 1–37
Shionoya S, Lu Y, Scanlon KJ (1986) Properties of amino acid transport systems in K562 cells sensitive and resistant tocis-diamminedichloroplatinum(II). Cancer Res 46: 3445–3448
Teicher BA, Holden SA, Kelley MJ (1987) Characterization of a human squamous carcinoma cell line resistant tocis-diamminedichloroplatinum(II). Cancer Res 47: 388–393
Vistica DT (1983) Cellular pharmacokinetics of the phenylalanine mustards. Pharmacol Ther 22: 379–405
Wallner KE, DeGregorio MW, Li GC (1986) Hyperthermic potentiation ofcis-diamminedichloroplatinum(II) cytotoxicity in Chinese hamster ovary cells resistant to the drug. Cancer Res 46: 6242–6245
Waud WR (1987) Differential uptake ofcis-diamminedichloroplatinum(II) by sensitive and resistant murine L1210 leukemia cells. Cancer Res 47: 6549–6555
Author information
Authors and Affiliations
Additional information
Supported by a grant from Bristol-Myers Co., grants CA-35309 and CA-23100 from the National Institutes of Health, and grants CH-368 and CH-417 from the American Cancer Society. This work was conducted in part by the Clayton Foundation for Research, California Division. Drs. Mann, Andrews, and Howell are Clayton Foundation investigators
Rights and permissions
About this article
Cite this article
Mann, S.C., Andrews, P.A. & Howell, S.B. Short-termcis-diamminedichloroplatinum(II) accumulation in sensitive and resistant human ovarian carcinoma cells. Cancer Chemother. Pharmacol. 25, 236–240 (1990). https://doi.org/10.1007/BF00684878
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00684878