Summary
The effect of tumor cell density on the cellular pharmacokinetics of doxorubicin (DXR) and cisplatin (CDDP) was studied using MOLT-3 human acute lymphoblastic leukemia cells. As determined by the MTT assay, the growth-inhibitory effect of DXR was approx. 40 times lower when cell density was increased from 106 to 108 cells/ml (positive inoculum effect), whereas little or no influence of cell density was observed in CDDP-induced cell-growth inhibition. As measured by high-performance liquid chromatography using a fluorescence detector, the cellular accumulation of DXR showed 6- and 18-fold decreases after 1 h incubation when the cells were concentrated from 106 to 107 and 108 cells/ml, respectively. Only at low cell density (106 cells/ml) did the amount of DXR in the cells increase with increasing exposure times of up to 6 h. The DXR concentration in the supernatant that was separated from a cell suspension showing a density of 108 cells/ml fell to 20% of that obtained at 106 cells/ml. The metabolites of DXR, including Adriamycinol and Adriamycinone, were not detectable in the cell extracts or supernatants at any cell density examined. In contrast, the cellular accumulation of CDDP calculated from the platinum concentration, which was measured with a flameless atomic absorption spectrophotometer, was essentially identical at all cell densities examined; moreover, extension of the exposure period resulted in a linear increase in the amount of CDDP in the cells. CDDP concentrations in the supernatants were equally retained, inrrespective of cell densities. These observations indicate that the positive inoculum effect shown in DXR-induced cell-growth inhibition results from the decreased cellular accumulation of the drug at high cell densities. We found no influence for cell density on the cellular accumulation of CDDP that might be relevant to the therapeutic potentiation of this drug at high tumor-cell density.
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Abbreviations
- DXR:
-
doxorubicin
- CDDP:
-
cisplatin,cis-diamminedichloroplatinum(II)
- HPLC:
-
high-performance liquid chromatography
- D-PBS:
-
Dulbecco's phosphate-buffered saline
- FBS:
-
fetal bovine serum
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
- ID50 :
-
drug concentration that produces inhibition of cell growth to 50% of control values
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This work was supported by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan (61 304 038) and by the Clinical Pathology Research Foundation of Japan
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Takemura, Y., Kobayashi, H., Miyachi, H. et al. The influence of tumor cell density on cellular accumulation of doxorubicin or cisplatin in vitro. Cancer Chemother. Pharmacol. 27, 417–422 (1991). https://doi.org/10.1007/BF00685154
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DOI: https://doi.org/10.1007/BF00685154