Skip to main content

Advertisement

SpringerLink
Log in

Short-Term Exposure of Cancer Cells to Micromolar Doses of Paclitaxel, with or without Hyperthermia, Induces Long-Term Inhibition of Cell Proliferation and Cell Death In Vitro

  • Published:
Annals of Surgical Oncology Aims and scope Submit manuscript

ABSTRACT

Background

During intraoperative hyperthermic intraperitoneal chemotherapy for primary or secondary peritoneal malignancies, tumor cells are exposed to high drug concentrations for a relatively short period of time. We investigated in vitro the effect of paclitaxel and hyperthermia on cell proliferation, cell cycle kinetics and cell death under conditions resembling those during intraoperative hyperthermic intraperitoneal chemotherapy.

Methods

Human breast MCF-7, ovarian SKOV-3 and hepatocarcinoma HEpG2 cells were exposed to 10 and 20 μM paclitaxel at 37, 41.5 or 43°C for 2 h. Cell proliferation, cell cycle kinetics, necrosis and apoptosis were evaluated.

Results

Hyperthermia exerted a cytostatic effect to all cell lines and at 43°C a cytotoxic effect on MCF-7 cells. MCF-7 and SKOV-3 cells treated under normothermic conditions with paclitaxel were arrested at G2/M or M phase for at least 3 days. Most of MCF-7 cells and approximately half of SKOV-3 cells were in interphase and became multinucleated without properly completing cytokinesis. Hyperthermia at 41.5°C altered cell cycle distribution and affected the paclitaxel-related effect on cell cycle kinetics of MCF-7 and SKOV-3 cells. Analysis of the mode of cell death showed that cell necrosis prevailed over apoptosis. Hyperthermia at 43°C increased paclitaxel-mediated cytotoxicity in MCF-7 cells and to a lesser extent in SKOV-3 and HEpG2 cells.

Conclusions

Short-time treatment of carcinoma cells with high (micromolar) concentrations of paclitaxel in normothermic and hyperthermic conditions is highly efficient for cell growth arrest and could be of clinical relevance in locoregional chemotherapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

FIG. 1.
FIG. 2.
FIG. 3.
FIG. 4.

Similar content being viewed by others

REFERENCES

  1. Markman M. Intraperitoneal antineoplastic drug delivery: rationale and results. Lancet Oncol 2004; 4:277–83

    Article  CAS  Google Scholar 

  2. Hofstra LS, de Vries EGE, Mulder NH, Willemse PHB. Intraperitoneal chemotherapy in ovarian cancer. Cancer Treat Rev 2000; 26:133–43

    Article  PubMed  CAS  Google Scholar 

  3. de Bree E, Rosing H, Michalakis J, et al. Intraperitoneal chemotherapy with taxanes for ovarian cancer with peritoneal dissemination. Eur J Surg Oncol 2006; 32:666–70

    Google Scholar 

  4. Mohamed F, Sugarbaker PH. Intraperitoneal taxanes. Surg Oncol Clin N Am 2003; 12:825–33

    Article  PubMed  Google Scholar 

  5. Sugarbaker PH, Welch LS, Mohamed F, Glehen O. A review of peritoneal mesothelioma at the Washington Cancer Institute. Surg Oncol Clin N Am 2003; 12:605–21

    Article  PubMed  Google Scholar 

  6. Ohashi N, Kodera Y, Nakanishi H, et al. Efficacy of intraperitoneal chemotherapy with paclitaxel targeting peritoneal micrometastases as revealed by GFP-tagged human gastric cancer cell lines in nude mice. Int J Oncol 2005; 27:637–44

    PubMed  CAS  Google Scholar 

  7. Feldman AL, Libutti SK, Pingpank JF, et al. Analysis of factors associated with outcome in patients with malignant peritoneal mesothelioma undergoing surgical debulking and intraperitoneal chemotherapy. J Clin Oncol 2003; 21:4560–7

    Article  PubMed  CAS  Google Scholar 

  8. Stewart JH IV, Shen P, Levine EA. Intraperitoneal hyperthermic chemotherapy for peritoneal surface malignancy: current status and future directions. Ann Surg Oncol 2005; 12:765–77

    Article  PubMed  Google Scholar 

  9. Witkamp AJ, de Bree E, Van Goethem R, Zoetmulder FAN. Rationale and techniques of intra-operative hyperthermic intraperitoneal chemotherapy. Cancer Treat Rev 2001; 27:365–74

    Article  PubMed  CAS  Google Scholar 

  10. Sticca RP, Dach BW. Rationale for hyperthermia with intraoperative intraperitoneal chemotherapy agents. Surg Oncol Clin N Am 2003; 12:689–701

    Article  PubMed  Google Scholar 

  11. Mohamed F, Marchettini P, Stuart OA, Urano M, Sugarbaker PH. Thermal enhancement of new chemotherapeutic agents at moderate hyperthermia. Ann Surg Oncol 2003; 10:463–8

    Article  PubMed  Google Scholar 

  12. Cividalli A, Cruciani G, Livdi E, Pasqualetti P, Tirindelli Danesi D. Hyperthermia enhances the response of paclitaxel and radiation in a mouse adenocarcinoma. Int J Radiat Oncol Biol Phys 1999; 44:407–12

    Article  PubMed  CAS  Google Scholar 

  13. Leal BZ, Meltz ML, Mohan N, Kuhn J, Prihoda TJ, Herman TS. Interaction of hyperthermia with Taxol in human MCF-7 breast adenocarcinoma cells. Int J Hyperthermia 1999; 15:225–36

    Article  PubMed  CAS  Google Scholar 

  14. Michalakis J, Georgatos SD, Romanos J, Koutala H, Georgoulias V, Tsiftsis D, Theodoropoulos PA. Micromolar taxol, with or without hyperthermia, induces mitotic catastrophe and cell necrosis in HeLa cells. Cancer Chemother Pharmacol 2005; 56:615–22

    Article  PubMed  CAS  Google Scholar 

  15. Othman T, Goto S, Lee JB, Taimura A, Matsumoto T, Kosaka M. Hyperthermic enhancement of the apoptotic and antiproliferative activities of paclitaxel. Pharmacology 2001; 62:208–12

    Article  PubMed  CAS  Google Scholar 

  16. van Bree C, Savonije JH, Franken NA, Haveman J, Bakker PJ. The effect of p53-function on the sensitivity to paclitaxel with or without hyperthermia in human colorectal carcinoma cells. Int J Oncol 2000; 16:739–44

    PubMed  Google Scholar 

  17. Rietbroek RC, Katschinski DM, Reijers MH, et al. Lack of thermal enhancement for taxanes in vitro. Int J Hyperthermia 1997; 13:525–33

    Article  PubMed  CAS  Google Scholar 

  18. Knox JD, Mitchel RE, Brown DL. Effects of taxol and taxol/hyperthermia treatments on the functional polarization of cytotoxic T lymphocytes. Cell Motil Cytoskeleton 1993; 24:129–38

    Article  PubMed  CAS  Google Scholar 

  19. Sharma D, Chelvi TP, Kaur J, Ralhan R. Thermosensitive liposomal taxol formulation: heatmediated targeted drug delivery in murine melanoma. Melanoma Res 1998; 8:240–4

    Article  PubMed  CAS  Google Scholar 

  20. Markman M, Kennedy A, Webster K, Kulp B, Peterson G, Belinson J. Carboplatin plus paclitaxel in the treatment of gynecologic malignancies: the Cleveland Clinic experience. Semin Oncol 1997; 24(5 Suppl 15):S15–26–9

    Google Scholar 

  21. de Bree E, Romanos J, Michalakis J, Relakis K, Georgoulias V, Melissas J, Tsiftsis DD. Intraoperative hyperthermic intraperitoneal chemotherapy with docetaxel as second-line treatment for peritoneal carcinomatosis of gynaecological origin. Anticancer Res 2003; 23:3019–27

    PubMed  Google Scholar 

  22. de Bree E, Rosing H, Beijnen JH, Romanos J, Michalakis J, Georgoulias V, Tsiftsis DD. Pharmacokinetic study of docetaxel in intraoperative hyperthermic i.p. chemotherapy for ovarian cancer. Anticancer Drugs 2003; 14:103–10

    Article  PubMed  Google Scholar 

  23. Rowinsky EK, Donehower RC. Paclitaxel (taxol). N Engl J Med 1995; 332:1004–14

    Article  PubMed  CAS  Google Scholar 

  24. Arnal I, Wade RH. How does taxol stabilize microtubules? Curr Biol 1995; 5:900–8

    Article  PubMed  CAS  Google Scholar 

  25. Mickey B, Howard J. Rigidity of microtubules is increased by stabilizing agents. J Cell Biol 1995; 130:909–17

    Article  PubMed  CAS  Google Scholar 

  26. 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 1993; 90:9552–6

    Article  PubMed  CAS  Google Scholar 

  27. Danesi R, Figg WD, Reed E, Myers CE. Paclitaxel (taxol) inhibits protein isoprenylation and induces apoptosis in PC-3 human prostate cancer cells. Mol Pharmacol 1995; 47:1106–11

    PubMed  CAS  Google Scholar 

  28. Theodoropoulos PA, Polioudaki H, Kostaki O, Derdas SP, Georgoulias V, Dargemont C, Georgatos SD. Taxol affects nuclear lamina and pore complex organization and inhibits import of karyophilic proteins into the cell nucleus. Cancer Res 1999; 59:4625–33

    PubMed  CAS  Google Scholar 

  29. Jordan MA, Wendell K, Gardiner S, Derry WB, Copp H, Wilson L. Mitotic block induced in HeLa cells by low concentrations of paclitaxel (Taxol) results in abnormal mitotic exit and apoptotic cell death. Cancer Res 1996; 56:816–25

    PubMed  CAS  Google Scholar 

  30. Bogdan C, Ding A. Taxol, a microtubule-stabilizing antineoplastic agent, induces expression of tumor necrosis factor alpha and interleukin-1 in macrophages. J Leukoc Biol 1992; 52:119–21

    PubMed  CAS  Google Scholar 

  31. Ding AH, Porteu F, Sanchez E, Nathan CF. Shared actions of endotoxin and taxol on TNF receptors and TNF release. Science 1990; 248:370–2

    Article  PubMed  CAS  Google Scholar 

  32. Lee LF, Schuerer-Maly CC, Lofquist AK, et al. Taxol-dependent transcriptional activation of IL-8 expression in a subset of human ovarian cancer. Cancer Res 1996; 56:1303–8

    PubMed  CAS  Google Scholar 

  33. Moos PJ, Fitzpatrick FA. Taxane-mediated gene induction is independent of microtubule stabilization: induction of transcription regulators and enzymes that modulate inflammation and apoptosis. Proc Natl Acad Sci USA 1998; 95:3896–901

    Article  PubMed  CAS  Google Scholar 

  34. Chan TA, Hermeking H, Lengauer C, Kinzler KW, Vogelstein B. 14-3-3Sigma is required to prevent mitotic catastrophe after DNA damage. Nature 1999; 401:616–20

    Article  PubMed  CAS  Google Scholar 

  35. Erenpreisa JE, Ivanov A, Dekena G, et al. Arrest in metaphase and anatomy of mitotic catastrophe: mild heat shock in two human osteosarcoma cell lines. Cell Biol Int 2000; 24:61–70

    Article  PubMed  CAS  Google Scholar 

  36. Merlin JL, Bour-Dill C, Marchal S, Bastien L, Gramain MP. Resistance to paclitaxel induces time-delayed multinucleation and DNA fragmentation into large fragments in MCF-7 human breast adenocarcinoma cells. Anticancer Drugs 2000; 11:295–302

    Article  PubMed  CAS  Google Scholar 

  37. Panvichian R, Orth K, Day ML, Day KC, Pilat MJ, Pienta KJ. Paclitaxel-associated multimininucleation is permitted by the inhibition of caspase activation: a potential early step in drug resistance. Cancer Res 1998; 58:4667–72

    PubMed  CAS  Google Scholar 

  38. Torres K, Horwitz SB. Mechanisms of Taxol-induced cell death are concentration dependent. Cancer Res 1998; 58:3620–6

    PubMed  CAS  Google Scholar 

  39. Blajeski AL, Kottke TJ, Kaufmann SH. A multistep model for paclitaxel-induced apoptosis in human breast cancer cell lines. Exp Cell Res 2001; 270:277–88

    Article  PubMed  CAS  Google Scholar 

  40. Yeung TK, Germond C, Chen X, Wang Z. The mode of action of taxol: apoptosis at low concentration and necrosis at high concentration. Biochem Biophys Res Commun 1999; 263:398–404

    Article  PubMed  CAS  Google Scholar 

  41. Chen JG, Yang CP, Cammer M, Horwitz SB. Gene expression and mitotic exit induced by microtubule-stabilizing drugs. Cancer Res 2003; 63:7891–9

    PubMed  CAS  Google Scholar 

  42. Abal M, Souto AA, Amat-Guerri F, Acuna AU, Andreu JM, Barasoain I. Centrosome and spindle pole microtubules are main targets of a fluorescent taxoid inducing cell death. Cell Motil Cytoskeleton 2001; 49:1–15

    Article  PubMed  CAS  Google Scholar 

  43. Bucci M, Wente SR. In vivo dynamics of nuclear pore complexes in yeast. J Cell Biol 1997; 136:1185–99

    Article  PubMed  CAS  Google Scholar 

Download references

ACKNOWLEDGMENTS

This work was supported partly by PENED 2001, 01EΔ376 from the Greek Secretariat of Research and Technology (PAT) and the Cretan Association for Biomedical Research (PAT and SDG).

Author information

Authors and Affiliations

  1. Department of Biochemistry, School of Medicine, University of Crete, P.O. Box 2208, 71003, Heraklion, Greece

    John Michalakis, Hara Polioudaki & Panayiotis A. Theodoropoulos

  2. Department of Surgical Oncology, School of Medicine, University of Crete, 71003, Heraklion, Greece

    John Michalakis, Eelco de Bree, John Romanos & Dimitris D. Tsiftsis

  3. Laboratory of Biology, School of Medicine, University of Ioannina, 45110, Ioannina, Greece

    Spyros D. Georgatos

  4. Department of Medical Oncology, School of Medicine, University of Crete, 71003, Heraklion, Greece

    Vassilis Georgoulias

Authors
  1. John Michalakis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Spyros D. Georgatos
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eelco de Bree
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hara Polioudaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. John Romanos
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Vassilis Georgoulias
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dimitris D. Tsiftsis
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Panayiotis A. Theodoropoulos
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Panayiotis A. Theodoropoulos.

Additional information

Authors Contribution:

JM: Acquisition, analysis and interpretation of data, drafting the manuscript; SDG: Design of the study, interpretation of data and revising the manuscript; EdB: Interpretation of data, drafting and revising the manuscript; HP: Acquisition of data and drafting the manuscript; JR and DDT: Conception of the study and revising the manuscript; VG: Interpretation of data and revising the manuscript; PAT: Conception and design of the study, analysis and interpretation of data, drafting and revising the manuscript. All authors have given final approval of the version to be published.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Michalakis, J., Georgatos, S.D., de Bree, E. et al. Short-Term Exposure of Cancer Cells to Micromolar Doses of Paclitaxel, with or without Hyperthermia, Induces Long-Term Inhibition of Cell Proliferation and Cell Death In Vitro. Ann Surg Oncol 14, 1220–1228 (2007). https://doi.org/10.1245/s10434-006-9305-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1245/s10434-006-9305-4

Keywords

Search

Navigation