Summary
2′,2′-Difluorodeoxycytidine (gemcitabine, dFdC) and cis-diammine-dichloroplatinum (cisplatin, CDDP) are active agents against ovarian cancer and non-small-cell lung cancer (NSCLC). CDDP acts by formation of platinum (Pt)–DNA adducts; dFdC by dFdCTP incorporation into DNA, subsequently leading to inhibition of exonuclease and DNA repair. Previously, synergism between both compounds was found in several human and murine cancer cell lines when cells were treated with these drugs in a constant ratio. In the present study we used different combinations of both drugs (one drug at its IC25 and the other in a concentration range) in the human ovarian cancer cell line A2780, its CDDP-resistant variant ADDP, its dFdC-resistant variant AG6000 and two NSCLC cell lines, H322 (human) and Lewis lung (LL) (murine). Cells were exposed for 4, 24 and 72 h with a total culture time of 96 h, and possible synergism was evaluated by median drug effect analysis by calculating a combination index (CI; CI < 1 indicates synergism). With CDDP at its IC25, the average CIs calculated at the IC50, IC75 IC90 and IC95 after 4, 24 and 72 h of exposure were < 1 for all cell lines, indicating synergism, except for the CI after 4 h exposure in the LL cell line which showed an additive effect. With dFdC at its IC25, the CIs for the combination with CDDP after 24 h were < 1 in all cell lines, except for the Cls after 4 h exposure in the LL and H322 cell lines which showed an additive effect. At 72 h exposure all Cls were < 1. CDDP did not significantly affect dFdCTP accumulation in all cell lines. CDDP increased dFdC incorporation into both DNA and RNA of the A2780 cell lines 33- and 79-fold (P < 0.01) respectively, and tended to increase the dFdC incorporation into RNA in all cell lines. In the AG6000 and LL cell lines, CDDP and dFdC induced > 25% more DNA strand breaks (DSB) than each drug alone; however, in the other cell lines no effect, or even a decrease in DSB, was observed. dFdC increased the cellular Pt accumulation after 24 h incubation only in the ADDP cell line. However, dFdC did enhance the Pt–DNA adduct formation in the A2780, AG6000, ADDP and LL cell lines (1.6-, 1.4-, 2.9- and 1.6-fold respectively). This increase in Pt–DNA adduct formation seems to be related to the incorporation of dFdC into DNA (r = 0.91). No increase in DNA platination was found in the H322 cell line. dFdC only increased Pt–DNA adduct retention in the A2780 and LL cell lines, but decreased the Pt–DNA adduct retention in the AG6000 cell line. In conclusion, the synergism between dFdC and CDDP appears to be mainly due to an increase in Pt–DNA adduct formation possibly related to changes in DNA due to dFdC incorporation into DNA.
Article PDF
Similar content being viewed by others
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Abbruzzese, J. L. & Frost, P. (1992). Studies on the mechanism of the synergistic interaction between 2′-deoxy-5-azacytidine and cisplatin. Cancer Chem Pharmacol 30: 31–36.
Abbruzzese, J. L., Grunawald, R., Weeks, E. A., Gravel, D., Adams, T., Nowak, B., Mineishi, S., Tarassoff, P., Satterlee, W., Raber, M. N. & Plunkett, W. (1991). A phase I clinical, plasma, and cellular pharmacology study of gemcitabine. J Clin Oncol 9: 491–498.
Abratt, R. P., Bezwoda, W. R., Goedhals, L. & Hacking, D. J. (1997). Weekly gemcitabine with monthly cisplatin: effective chemotherapy for advanced non-small cell lung cancer. J Clin Oncol 15: 744–749.
Bergman, A. M., Ruiz van Haperen, V. W. T., Veerman, G., Kuiper, C. M. & Peters, G. J. (1996). Interaction between cisplatin and gemcitabine in vitro. Clin Cancer Res 2: 521–530.
Birnboim, H. C. & Jevcak, J. J. (1981). Fluorometric method for rapid detection of DNA strand breaks in human white blood cells produced by low doses of radiation. Cancer Res 41: 1889–1892.
Braakhuis, B. J. M., Ruiz van Haperen, V. W. T., Welters, M. J. P. & Peters, G. J. (1995). Schedule-dependent therapeutic efficacy of the combination of gemcitabine and cisplatin in head and neck cancer xenografts. Eur J Cancer 31A: 1335–1340.
Chiu, C. S. M., Chan, A. K. & Wright, J. A. (1992). Inhibition of mammalian ribonucleotide reductase by cis-diamminedichloroplatinum(II). Biochem Cell Biol 70: 1332–1338.
Chou, T-C & Hayball, M. P. (1996). CalcuSyn, Windows Software for Dose Effect Analysis. Biosoft: Cambridge
Chou, T-C & Talalay, P. (1983). Quantitative analysis of dose-effect relationship: the combined effects of multiple drugs on enzyme inhibitors. In Advances in Enzyme Regulation, G. Weber (ed), pp. 27–55. Pergamon Press: New York
Chou, T-C, Motzer, R. J., Tong, Y. & Bosl, G. J. (1994). Computerized quantitation of synergism and antagonism of taxol, topotecan, and cisplatin against human teratocarcinoma cell growth: a rational approach to clinical protocol design. J Natl Cancer Inst 86: 1517–1524.
Crino, L., Scagliotto, G., Marangolo, M., Figoli, F., Clerici, M., DeMarinis, F., Salvati, F., Cruciani, G., Dagliotti, L., Pucci, F., Paccagnella, A., Adamao, V., Altavilla, G., Incoronato, P., Tripetti, M., Mosconi, A. M., Santucci, A., Sorbolini, S., Oliva, C. & Tonato, M. (1997). Cisplatin-gemcitabine combination in advanced non-small cell lung cancer: a phase II study. J Clin Oncol 15: 297–303.
Ellerhorst, J. A., Frost, P., Abbruzzese, J. L., Newman, R. A. & Chernajovsky, Y. (1993). 2′-deoxy-5-azacytidine increases binding of cisplatin to DNA by a mechanism independent of DNA hypomethylation. Br J Cancer 67: 209–215.
Freeman, K. B., Anliker, S., Hamilton, M., Osborne, D., Dhahir, P. H., Nelson, R. & Allerheiligen, S. R. B. (1995). Validated assays for the determination of gemcitabine in human plasma and urine using high-performance liquid-chromatography with ultraviolet detection. J Chromatogr Biomed Appl 665: 171–181.
Heinemann, V., Hertel, L. W., Grindey, G. B. & Plunkett, W. (1988). Comparison of the cellular pharmacokinetics and toxicity of 2′,2′-difluorodeoxycytidine and 1-β-D-arabinofuranosylcytosine. Cancer Res 48: 4024–4031.
Hertel, L. W., Kroin, J. S., Misner, J. W. & Tustin, J. M. (1988). Synthesis of 2′-deoxy-2′,2′-difluoro-D-ribose and 2′-deoxy-2′,2′-difluoro-D-ribofuranosyl nucleosides. J Org Chem 53: 2406–2409.
Huang, P., Chubb, S., Hertel, L. W., Grindey, G. B. & Plunkett, W. (1991). Action of 2′,2′-difluorodeoxycytidine on DNA synthesis. Cancer Res 51: 6110–6117.
Krakowski, I., Petit, T., Kayitalire, L., Weber, B., Beaudouin, M., Canon, J. L., Janssens, J., Martin, C. & Belpomme, D. (1998). Gemcitabine (Gemzar) in combination with cisplatin (CP) in advanced ovarian cancers (AOC): a phase II study. Proc Am Soc Clin Oncol 17: 356a (abstract 1373)
Lu, Y., Han, J. & Scanlon, K. J. (1988). Biochemical and molecular properties of cisplatin-resistant A2780 cells grown in folinic acid. J Biol Chem 263: 4891–4894.
Monks, A., Scudiero, D., Skehan, P., Shoemaker, R., Paull, K., Vistica, D., Hose, C., Langley, J., Cronise, P., Vaigro-Wolff, A., Gray-Goodrich, M., Campbell, H., Mayo, J. & Boyd, M. (1991). Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines. J Natl Cancer Inst 83: 761–766.
Nogué, M., Cirera, L., Arcusa, M., Tusquets, I., Batiste-Alentorn, E., Font, A. & Boto, B. (1998). Gemcitabine combined with cisplatin first line: a phase II study in patients with advanced epithelial ovarian cancer. Proc Am Soc Clin Oncol 17: 357a (abstract 1377)
Parker, R. J., Gill, I., Tarone, R., Vionnet, J., Grunberg, S., Muggia, F. & Reed, E. (1991). Platinum DNA-damage in leucocyte DNA of patients receiving cisplatin and carboplatin chemotherapy, measured by atomic absorption spectrometry. Carcinogenesis 12: 1253–1258.
Peters, G. J., Laurensse, E., Leyva, A. & Pinedo, H. M. (1987). Purine nucleosides as cell-specific modulators of 5-fluorouracil metabolism and cytotoxicity. Eur J Cancer Clin Oncol 23: 1869–1881.
Peters, G. J., Wets, M., Keepers, YPAM, Oskam, R., Van Ark-Otte, J., Noordhuis, P., Smid, K. & Pinedo, H. M. (1993a). Transformation of mouse fibroblasts with the oncogenes H-ras or trk is associated with pronounced changes in drug sensitivity and metabolism. Int J Cancer 54: 450–455.
Peters, G. J., Schornagel, J. H. & Milano, G. A. (1993b). Clinical pharmacokinetics of anti-metabolites. Cancer Surveys 17: 123–156.
Ruiz van Haperen, V. W. T., Veerman, G., Vermorken, J. B. & Peters, G. J. (1993). 2′,2′-Difluoro-deoxocytidine (Gemcitabine) incorporation into RNA and DNA of tumour cell lines. Biochem Pharmacol 46: 762–766.
Ruiz van Haperen, V. W. T., Veerman, G., Eriksson, S., Boven, E., Stegmann, A. P. A., Hermsen, M., Vermorken, J. B., Pinedo, H. M. & Peters, G. J. (1994a). Development and characterization of a 2′,2′-difluorodeoxycytidine-resistant variant of the human ovarian cancer cell line A2780. Cancer Res 54: 4138–4143.
Ruiz van Haperen, V. W. T., Veerman, G., Boven, E., Noordhuis, P., Vermorken, J. B. & Peters, G. J. (1994b). Schedule-dependence of sensitivity to 2′,2′-difluorodeoxycytidine (Gemcitabine) in relation to accumulation and retention of its triphosphate in solid tumor cell lines and solid tumors. Biochem Pharmacol 48: 1327–1339.
Ruiz van Haperen, V. W. T., Veerman, G., Vermorken, J. B., Pinedo, H. M. & Peters, G. J. (1996). Regulation of deoxycytidine kinase from solid tumor cell lines by CTP and UTP. Biochem Pharmacol 51: 911–918.
Scanlon, K. J., Kashai-Sabet, M., Tone, T. & Funato, T. (1991). Cisplatin resistance in human cancers. Pharmacol Ther 52: 385–406.
Steward, W. P., Dunlop, D. J., Dabouis, G., Lacroix, H. & Talbot, D. (1996). Phase I/II study of gemcitabine and cisplatin in non-small cell lung cancer: preliminary results. Semin Oncol 5: 43–47.
Sundquist, W. I. & Lippard, S. J. (1990). The coordination chemistry of platinum anticancer drugs and related compounds with DNA. Coord Chem Rev 100: 293–322.
Terheggen, PMAB, Emondt, J. Y., Floot, B. G. J., Dijkman, R., Schrier, P. I., Den Engelse, L. & Begg, A. C. (1990). Correlation between cell killing by cis-diamminedichloroplatinum(II) in six mammalian cell lines and binding of a cis-diamminedichloroplatinum(II)-DNA antiserum. Cancer Res 50: 3556–3561.
Van der Vijgh, W. J. F. (1991). Clinical pharmacology of carboplatin. Clin Pharmacokin 21: 242–261.
Van der Wilt, C. L., Smid, K., Noordhuis, P., Aherne, G. W. & Peters, G. J. (1997). Biochemical mechanisms of interferon modulation of 5-fluorouracil activity in colon cancer cells. Eur J Cancer 33: 471–478.
Van der Wilt, C. L., Visser, G. W. M., Braakhuis, B. J. M., Wedzinga, R., Noordhuis, P., Smid, K. & Peters, G. J. (1993). In vitro antitumour activity of cis- and trans-5-fluoro-5,6-dihydro-6-alkoxy-uracils. Br J Cancer 68: 702–707.
Van Moorsel, C. J. A., Peters, G. J. & Pinedo, H. M. (1997). Gemcitabine: future prospects of single-agent and combination studies. Oncologist 2: 127–134.
Van Moorsel, C. J. A., Pinedo, H. M., Veerman, G., Vermorken, J. B., Postmus, P. E. & Peters, G. J. (1999). Scheduling of gemcitabine and cisplatin in Lewis Lung tumour bearing mice. Eur J Cancer. In press
Vermorken, J. B., Van der Vijgh, W. J. F., Klein, I., Gall, H. E., Van Groeningen, C. J. & Pinedo, H. M. (1984). Pharmacokinetics of free and total platinum species after rapid and prolonged infusions of cisplatin. Clin Pharmacol Ther 39: 136–144.
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Moorsel, C., Pinedo, H., Veerman, G. et al. Mechanisms of synergism between cisplatin and gemcitabine in ovarian and non-small-cell lung cancer cell lines. Br J Cancer 80, 981–990 (1999). https://doi.org/10.1038/sj.bjc.6690452
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6690452
- Springer Nature Limited