Abstract
Paclitaxel (Taxol®) kills tumor cells by inducing both cellular necrosis and apoptosis. A major impediment to paclitaxel cytotoxicity is the establishment of multidrug resistance whereby exposure to one chemotherapeutic agent results in cross-resistance to a wide variety of other drugs. For example, selection of MCF-7 breast cancer cells for resistance to doxorubicin (MCF-7ADR cells) results in cross-resistance to paclitaxel. This appears to involve the overexpression of the drug transporter P-glycoprotein which can efflux both drugs from tumor cells. However, MCF-7ADR cells possess a deletion mutation in p53 and have considerably reduced levels of the Fas receptor, Fas ligand, caspase-2, caspase-6, and caspase-8, suggesting that paclitaxel resistance may also stem from a bona fide block in paclitaxel-induced apoptosis in these cells. To address this issue, we examined the ability of the P-glycoprotein inhibitor valspodar to restore paclitaxel accumulation, paclitaxel cytotoxicity, and paclitaxel-induced apoptosis. Compared to drug sensitive MCF-7 cells, MCF-7ADR cells accumulated >6-fold less paclitaxel, were approximately 100-fold more resistant to killing by the drug, and were highly resistant to paclitaxel-induced apoptosis. In contrast, MCF-7ADR cells pretreated with valspodar were indistinguishable from drug-sensitive cells in their ability to accumulate paclitaxel, in their chemosensitivity to the drug, and in their ability to undergo paclitaxel-induced apoptosis. Valspodar, by itself, did not affect these parameters. This suggests that the enhancement of paclitaxel toxicity in MCF-7ADR cells involves a restoration of apoptosis and not solely through enhanced drug-induced necrosis. Morever, it appears that changes in the levels/activity of p53, the Fas receptor, Fas ligand, caspase-2, caspase-6, or caspase-8 activity have little effect on paclitaxel-induced cytotoxicity and apoptosis in human breast cancer cells.
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Seidman AD, Reichman BS, Crown JPA, Curie V, Hakes TB, Hudis CA, Gilewski TA, Baselga J, Forsythe P, Lepore J, Marks L, Fain K, Southrada M, Onetta N, Arbuck S, Norton I: Paclitaxel as second and subsequent therapy for breast cancer. J Clin Onc 13: 1152–1159, 1995
Crown J: Optimising treatment outcomes: a review of current management strategies in first-line chemotherapy of metastatic breast cancer. Eur J Cancer 33: S15–S19, 1997 (Suppl 7)
Wiseman LR, Spencer CM: Paclitaxel. An update of its use in the treatment of metastatic breast cancer and ovarian and other gynaecological cancers. Drugs Aging 12: 305–334, 1998
Wani MC, Taylor HL, Wall ME, Coggan P, McPhail AT: Plant antitumor agents V1. The isolation and structure of taxol, a novel antileukemic and antitumor agent from taxus brevifolia. J Am Chem Soc 93: 2325–2327, 1971
Schiff PB, Fant J, Horwitz SB: Promotion of microtubule assembly in vitro by taxol. Nature 277: 665–667, 1979
De Brabander M, Geuens G, Nuydens R, Willebrords R, De Mey J: Taxol induces the assembly of free microtubules in living cells and blocks the organizing capacity of the centrosomes and kinetochores. Proc Natl Acad Sci USA 78: 5608–5612, 1981
Jordan MA, Toso RJ, Thrower D, Wilson, L: Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations. Proc Natl Acad Sci USA 90: 9552–9556, 1993
Bhalla K, Ibrado AM, Tourkina F, Tang C, Mahoney ME, Huang Y: Taxol induces internucleosomal DNA fragmentation associated with programmed cell death in human myeloid leukemia cells. Leukemia 7: 563–688, 1993
Allen RT, Hunter WJ III, Agrawal DK: Morphological and biochemical characterization and analysis of apoptosis. J Pharmacol Toxicol Meth 37: 215–228, 1997
Janicke RU, Sprengart ML, Wati MR, Porter AG: Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem 273: 9357–9360, 1998
Saunders DE, Lawrence WD, Christensen C, Wappler NL, Ruan H, Deppe G: Paclitaxel-induced apoptosis in MCF-7 breast cancer cells. Int J Cancer 70: 214–220, 1997
Chen G, Teicher BA, Frei E III: Differential interactions of P-glycoprotein inhibitor thaliblastine with adriamycin, etoposide, taxol and anthrapyrazole C1941 in sensitive and multidrug-resistant human MCF-7 breast cancer cells. Anticancer Res 16: 3499–3505, 1996
Juliano RL, Ling V: A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta 455: 152–162, 1976
Beck WT, Mueller TJ, Tanzer LR: Altered surface membrane glycoproteins in Vinca alkaloid-resistant human leukemic lymphoblasts. Cancer Res 39: 2070–2076, 1979
Cole SPC, Bhardwaj G, Gerlach JH, Mackie JE, Grant CE, Almquist KC, Stewart AJ, Kurz EU, Duncan AMV, Deeley RG: Overexpression of a transporter gene in a multidrugresistant human lung cancer cell line. Science 258: 1650–1654, 1992
Grant CE, Valdimarsson G, Hipfner DR, Almquist KC, Cole SPC, Deeley RG: Overexpression of multidrug resistanceassociated protein (MRP) increases resistance to natural product drugs. Cancer Res 54: 357–361, 1994
Harris AL, Hochhauser D: Mechanisms of multidrug resistance in cancer treatment. Acta Oncol 31: 205–213, 1992
Fry AM, Chresta CM, Davies SM, Walker MC, Harris AL, Hartley JA, Masters JR, Hickson, ID: Relationship between tomoisomerase II level and chemosensitivity in human tumor cell lines. Cancer Res 15: 6592–6595, 1991
Giaccone G, Gazdar AF, Beck H, Zunino F, Capranico G: The multidrug sensitivity phenotype of human lung cancer cells associated with topoisomerase II expression. Cancer Res 52: 1666–1674, 1992
Friesen C, Fulda S, Debatin KM: Deficient activation of the CD9S (APO-1/Fas) system in drug-resistant cells. Leukemia 11: 1833–1841, 1997
Mechetner E, Kyshtoobayeva A, Zonis S, Kim H, Stroup R, Garcia R, Parker RJ, Fruehauf JP: Levels of multidrug resistance (MDR1) P-glycoprotein expression by human breast cancer correlate with in vitro resistance to taxol and doxorubicin. Clin Cancer Res 4: 389–398, 1998
Ogretmen B, Safa AR: Expression of the mutated p53 tumor supressor protein and its molecular and biochemical characterization in multidrug resistant MCF-7/Adr human breast cancer cells. Oncogene 14: 499–506, 1997
Cai Z, Stancou R, Korner M, Chouaib S: Impairment of Fas-antigen expression in adriamycin-resistant but not TNFresistant MCF7 tumor cells. Int J Cancer 68: 535–546, 1996
Jachez B, Nordmann R, Loor F: Restoration oftaxol sensitivity of multi-drug resistant cells by the cyclosporine SDZ-PSC833 and the cyclopeptolide SDZ280-446. J Natl Cancer Inst 85: 478–483, 1993
Arichnal-Mattheis A, Rzepka RW, Watanabe T, Kokob N, Itoh Y, Combates NJ, Bair KW, Cohen, D: Analysis of the interactions of SDZ-PSC83 3 ([30-keto-Bmt1]-Val2-Cyclosporine), a multidrug resistant modulator, with P-glycoprotein. Oncol Res 7: 603–610, 1995
Wang Z, Tung PS, Moran, MF: Association of P120 ras gap with endocytic components and colocalization with epidermal growth factor (EGF) receptor in response to EGF stimulation. Cell Growth Differ 7: 123–133, 1996
Hubbard AL, Wall AD, Ma M: Isolation of rat hepatocyte plasma membranes. I. Presence of the Three Major Domains. J Cell Biol 96: 217–229, 1983
Keating A, Toor P: Human long term bone marrow culture. In: Poclard JM (ed) Methods in Molecular Biology. Wiley-Liss, New York, Vol 5, 1990, p 331
Mercille S, Massie B: Induction of apoptosis in nutrientdeprived cultures of hybridoma and myeloma cells. Biotech Bioeng 44: 1140–1154, 1994
Fadok VA, Voelker DR, Campbell PA, Cohen JJ, Bratton DL, Henson PM: Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J Immunol 148: 2207–2216, 1992
Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MH: Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84: 1415–1420
Fairchild CR, Ivy SP, Kao-Shan CS, Whang-Peng J, Rosen N, Israel MA, Melera PW, Cowan KH, Goldsmith ME: Isolation of amplified and overexpressed DNA sequences from adriamycin-resistant human breast cancer cells. Cancer Res 47: 5141–5148, 1987
Cowan KH, Batist G, Tulpule A, Sinha BK, Myers CE: Similar biochemical changes associated with multidrug resistance in human breast cancer cells and carcinogen-induced resistance to xenobiotics in rats. Proc Natl Acad Sci USA 83: 9328–9332, 1986
Batist G, Tulpule A, Sinha BK, Katki AG, Myers CE, Cowan, KH: Overexpression of a novel anionic glutathione transferase in multidrug-resistant human breast cancer cells. J Biol Chem 261: 15544–15549, 1986
Doroshow JH: Prevention of doxorubicin-induced killing of MCF-7 human breast cancer cells by oxygen radical scavengers and iron chelating agents. Biochem Biophys Res Commun 135: 330–335, 1986
Davies R, Budworth J, Riley J, Snowden R, Gescher A, Gant TW: Regulation of P-glycoprotein 1 and 2 gene expression and protein activity in two MCF-7/Dox cell line subclones. Br J Cancer 73, 307–315, 1996
Yamashima T, Ohnishi T, Nakajima Y, Terasaki T, Tanaka M, Yamashita J, Sasaki T, Tsuji A: Uptake of drugs and expression of P-glycoprotein in the rat 9L glioma. Exp Brain Res 95: 41–50, 1993
Eischen CM, Kottke TJ, Martins LM, Basil GS, Tung JS, Eamshaw WC, Leibson PJ, Kaufmann SH: Comparison of apoptosis in wild-type and Fas-resistant cells: chemotherapyinduced apoptosis is not dependent on Fas/Fas ligand interactions. Blood 90: 935–943, 1997
Woods CM, Zhui J, McQueney A, Bollag D, Lazarides E: Taxol-induced mitotic block triggers rapid onset of a p53 independent apoptotic pathway. Molec Med 95: 1076–1151, 1995
Strobel T, Swanson L, Korsmeyer S, Cannistra SA: BAX enhances paclitaxel-induced apoptosis through a p53-independent pathway. Proc Natl Acad Sci USA 93: 14094–14099, 1996
Vasey PA, Jones NA, Jenkins S, Dive C, Brown R: Cisplatin, camptothecin, and taxol sensitivities of cells with p53-associated multidrug resistance. Mol Pharmacol 50: 1536–1540, 1996
Debernardis D, Sire EG, De Feudis P, Vikhanskaya F, Valenti M, Russo P, Parodi S, D'Incalci M, Broggini M: p53 status does not affect sensitivity of human ovarian cancer cell lines to paclitaxel. Cancer Res 57: 870–874, 1997
Jones NA, Turner J, McIlwrath AJ, Brown R, Dive C: Cisplatin-and paclitaxel-induced apoptosis of ovarian carcinoma cells and the relationship between bax and bak upregulation and the functional status of p53. Mol Pharmacol 53: 819–826, 1998
Zaffaroni N, Silvestrini R, Orlandi L, Bearzatto A, Gornati D, Villa R: Induction of apoptosis by taxol and cisplatin and effect on cell cycle-related proteins in cisplatin-sensitive and-resistant human ovarian cells. Br J Cancer 77: 1378–1385, 1998
Bachman CA, Bills DA, Majumdar SK: Evidence of p53-induced apoptosis in cancer cells exposed to taxol. In Vitro Cell Dev Biol Anim 34: 434–435, 1998
Torres K, Horwitz SB: Mechanisms oftaxol-induced cell death are concentration dependent. Cancer Res 58: 3620–3626, 1998
Roth W, Wagenknecht B, Grimmel C, Dichgans J, Weller M: Taxol-mediated augmentation of CD95 ligand-induced apoptosis of human malignant glioma cells: association with bcl-2 phosphorylation but neither activation of p53 nor G2/M cell cycle arrest. Br J Cancer 77: 404–411, 1998
Lanni JS, Lowe SW, Licitra EJ, Liu JO, Jacks T: p53-independent apoptosis induced by paclitaxel through an indirect mechanism. Proc Natl Acad Sci USA 94: 9679–9683, 1997
Rodi DJ, Janes RW, Sanganee HJ, Holton RA, Wallace BA, Makowski L: Screening of a library of phage-displayed peptides identifies human bcl-2 as a taxol-binding protein. J Mol Biol 285: 197–203, 1999
Thornberry NA, Lazebnik Y: Caspases: enemies within. Science 281: 1312–1316, 1998
Fan W: Possible mechanisms of paclitaxel-induced apoptosis. Biochem Pharmacol 57: 1215–1221, 1999
Milross CG, Mason KA, Hunter NR, Chung WK, Peters LJ, Milas L: Relationship of mitotic arrest and apoptosis to antitumor effect of paclitaxel. J Natl Cancer Inst 18: 1308–1314, 1996
Scudiero DA, Monks A, Sausville BA: Cell line designation change: multidrug-resistant cell line in the NCI anticancer screen. J Natl Cancer Inst 90: 862, 1998
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Chadderton, A., Villeneuve, D.J., Gluck, S. et al. Role of specific apoptotic pathways in the restoration of paclitaxel-induced apoptosis by valspodar in doxorubicin-resistant MCF-7 breast cancer cells. Breast Cancer Res Treat 59, 231–244 (2000). https://doi.org/10.1023/A:1006344200094
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DOI: https://doi.org/10.1023/A:1006344200094