Abstract
Purpose
In this study, the cytotoxic effects of paclitaxel (PAC) in normal and tumor cells were established, and the cytoprotective effects of amifostine (AMI) and cysteamine (CYS) against this cytotoxicity were examined.
Methods
Tumor cell lines used in this study were L-strain cells of mouse subcutaneous origin and human cervix carcinoma-derived HeLa cells. Mouse embryonic fibroblasts (MEFs) were used as the normal cell line. Results of the experiments were evaluated in terms of the mitotic index and the 3H-thymidine labeling index. PAC concentrations of 6 and 12 μg/ml were applied to the cells for 1–10 days either alone or in combination with 1 μg/ml of AMI and CYS.
Results
In terms of the above parameters, statistically significant effects were not seen in cultures of any of the cell lines treated with 1 μg/ml of AMI or CYS alone. In contrast, both concentrations of PAC caused increasing cytotoxic effects with increasing treatment time (P<0.001). The cytotoxic effect of PAC appeared as mitotic phase accumulation (G2/M blockage) and a subsequent decline in the synthesis phase. HeLa cells were very sensitive to PAC treatment, whereas MEF cells were quite resistant compared with tumor cells. In cells treated with combined drugs to investigate the cytoprotective effects of AMI and CYS on normal and tumor cell lines, PAC continued to show cytotoxic effects in tumor cells, but this effect was reduced in the normal cells.
Conclusions
AMI and CYS did not protect tumor cells against the cytotoxic effects of PAC, but protection was observed in normal cells. Furthermore, the protection provided by AMI was stronger than that provided by CYS.
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References
Acosta JC, Richard C, Delgado MD, Horita M, Rizzo MG, Fernandez-Luna JL, Leon J (2003) Amifostine impairs p53-mediated apoptosis of human myeloid leukemia cells. Mol Cancer Ther 2:893–900
Brenner B, Wasserman L, Beery E, Nordenberg J, Schechter J, Gutman H, Fenig E (2003) Variable cytotoxicity of amifostine in malignant and non-malignant cell lines. Oncol Rep 10(5):1609–1613
Castiglione F, Dalla Mola A, Porcile G (1999) Protection of normal tissues from radiation and cytotoxic therapy: the development of amifostine. Tumori 85(2):85–91
Davis W Jr, Ronai Z, Tew KD (2001) Cellular thiols and reactive oxygen species in drug-induced apoptosis. J Pharmacol Exp Ther 296:1–6
de Matos DG, Herrera C, Cortvrindt R, Smitz J, Van Soom A, Nogueria D, Pasqualini RS (2002) Cysteamine supplementation during in vitro maturation and embryo culture: a useful tool for increasing the efficiency of bovine in vitro embryo production. Mol Reprod Dev 62(2):203–209
Dorr RT (1996) Cytoprotective agents for anthracyclines. Semin Oncol 23(4 Suppl 8):23–34
Freshney RI (1991) Disaggregation of the tissue and primary culture. In: Culture of animal cells. A manual of basic technique, 2nd edn. Wiley Liss, New York, pp 107–126
Freshney RI (1991) Maintenance of the culture-cell lines. In: Culture of animal cells. A manual of basic technique, 2nd edn. Wiley Liss, New York, pp 127–136
Fulda S, Oster W, Berthold F (1997) Effects of WR-2721 (amifostine) and its metabolite WR-1065 on the antiproliferative activity of chemotherapeutic agents on neuroblastoma cells in vitro. Anticancer Drugs 8(1):34–41
Gelmon K, Eisenhauer E, Bryce C, Tolcher A, Mayer L, Tomlinson E, Zee B, Blackstein M, Tomiak E, Yau J, Batist G, Fisher B, Iglesias J (1999) Randomized phase II study of high-dose paclitaxel with or without amifostine in patients with metastatic breast cancer. J Clin Oncol 17(10):3038–3047
Gentsch JR, Hatfield JW (1984) Saturable attachment sites for type 3 mammalian reovirus on murine L cells and human HeLa cells. Virus Res 1(5):401–414
Lee EJ, Gerhold M, Palmer MW, Christen RD (2003) p53 protein regulates the effects of amifostine on apoptosis, cell cycle progression, and cytoprotection. Br J Cancer 88:754–759
Lopes NM, Adams EG, Pitts TW (1993) Cell kill kinetics and cell cycle effects of taxol on human and hamster ovarian cell lines. Cancer Chemother Pharmacol 32:235–242
Modig HG, Edgren M, Révész L (1971) Release of thiols from cellular mixed disulphides and its possible role in radiation protection. Int Radiat Biol 22(3):257–268
Ng TY, Ngan HY, Cheng DK, Wong LC (1999) The effect of amifostine on the in vitro cytotoxicity of chemotherapeutic agents in three epithelial ovarian carcinoma cell lines. Gynecol Oncol 75(2):194–197
Obata S, Yamaguchi Y, Miyamoto T (1990) Effects of etoposide (VP-16) on the survival and progression of cultured HeLa S3 cells through the cell cycle. Nippon Gan Chiryo Gakkai Shi 25(10):2484–2491
Özcan FG, Topčul MR, Yılmazer N, Rıdvanoğulları M (1997) Effect of epirubicin on 3H-thymidine labelling index in cultured L-strain cells. J Exp Clin Cancer Res 16(1):23–27
Paine GD, Taylor CW, Lopez MH, Johnson CS, Capizzi RL (1996) Effects of amifostine and paclitaxel on growth of human ovary carcinoma xenografts in the severe combined immune-deficiency mouse: preliminary results. Semin Oncol 23(4 Suppl 8):35–39
Purdie JW (1979) A comparative study of the radioprotective effects of cysteamine, WR-2721 and WR-1065 in cultured human cells. Radiat Res 77(2):303–311
Rowinsky EK, Onetto N, Canetta RM, Arbuck SG (1992) Taxol: the first of the taxanes, an important new class of antitumor agents. Semin Oncol 19:646–662
Santini V, Giles FJ (1999) The potential of amifostine: from cytoprotectant to therapeutic agent. Haematologica 84(11):1035–1042
Shannon JE (1972) The American Type Culture Collection. Registry of animal cell lines, vol 301. ATCC, Rockville, pp 881–2600
Shen H, Chen Z, Zilfou JT, Hopper E, Murphy M, Tew KD (2001) Binding of the aminothiol WR-1065 to transcription factors influences cellular response to anticancer drugs. Pharmacology 297(3):1067–1073
Spencer CM, Goa KL (1995) Amifostine: a review of its pharmacodynamic and pharmacokinetic properties, and therapeutic potential as a radioprotector and cytotoxic chemoprotector. Drugs 50(6):1001–1031
Takahashi M, Nagai T, Hamano S, Kuwayama M, Okamura N, Okano A (1993) Effect of thiol compounds on in vitro development and intracellular glutathione content of bovine embryos. Biol Reprod 49(2):228–232
Taylor CW, Wang LM, List AF, Fernandes D, Paine-Murrieta GD, Johnson CS, Capizzi RL (1997) Amifostine protects normal tissues from paclitaxel toxicity while cytotoxicity against tumor cells is maintained. Eur J Cancer 33(10):1693–1698
Wang Z, Yu R, Melmed S (2001) Mice lacking pituitary tumor transforming gene show testicular and splenic hypoplasia, thymic hyperplasia, thrombocytopenia, aberrant cell cycle progression, and premature centromere division. Mol Endocrinol 15(11):1870–1879
Zhang X, Lai PP, Taylor YC (1992) Differential radioprotection of cultured human diploid fibroblasts and fibrosarcoma cells by WR-1065. Int J Radiat Oncol Biol Phys 24:713–719
Acknowledgements
This work was supported by the Research Fund of The University of Istanbul, project no. T-560/240698. The author would like to thank Bristol Myers Squibb for kindly supplying paclitaxel (Taxol) for use in this work. The author is also grateful to Prof. Dr. Atilla Özalpan for his helpful suggestions and critical remarks.
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Özcan Arican, G. Cytoprotective effects of amifostine and cysteamine on cultured normal and tumor cells treated with paclitaxel in terms of mitotic index and 3H-thymidine labeling index. Cancer Chemother Pharmacol 56, 221–229 (2005). https://doi.org/10.1007/s00280-004-0954-3
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DOI: https://doi.org/10.1007/s00280-004-0954-3