Skip to main content
SpringerLink
Log in

P-glycoprotein expression does not change the apoptotic pathway induced by curcumin in HL-60 cells

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

One of the mechanisms responsible for the multidrug resistance (MDR) phenotype of cancer cells is overexpression of so-called ATP-dependent drug efflux proteins: the 170-kDa P-glycoprotein (P-gp) encoded by the MDR1 gene and the 190-kDa multidrug resistance-associated protein 1 encoded by the MRP1 gene. The purpose of the present study was to verify the hypothesis postulating that P-gp expression, apart from enabling drug efflux, confers on the cells resistance to apoptosis by inhibiting caspase-8 and caspase-3.

Materials and methods

Human HL-60 cells, either drug-sensitive or with the MDR phenotype caused by overexpression of P-gp (HL-60/Vinc) or MRP1 (HL-60/Adr), were treated with the natural dye curcumin at 50 μM or with UVC to induce apoptosis. Symptoms of cell death were assessed by morphological observation after Hoechst staining, DNA fragmentation was measured by flow cytometry and the TUNEL method, and caspase-8 and caspase-3 activation and cytochrome c release from mitochondria were measured by Western blotting.

Results

Curcumin induced cell death in HL-60 cells, both sensitive and with the MDR phenotype, which could be classified as caspase-3-dependent apoptosis, together with cytochrome c release, activation of caspase-3 and oligonucleosomal DNA fragmentation. No active caspase-8 was detected. Also UVC caused caspase-3 activation in both the sensitive and the MDR HL-60 cells.

Conclusions

Our findings show that there was no correlation between P-gp expression and resistance to caspase-3-dependent apoptosis induced by curcumin and UVC, at least in HL-60 cells. However, we cannot exclude the possibility of parallel P-gp expression and caspase-3 inhibition in some other cell lines, as cancer cells can acquire many different apoptosis-resistance mechanisms.

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. 3A, B
Fig. 4
Fig. 5A, B
Fig. 6

Similar content being viewed by others

References

  1. Bielak-Żmijewska A, Koronkiewicz M, Skierski J, Piwocka K, Radziszewska E, Sikora E (2000) Effect of curcumin on the apoptosis of rodent and human nonproliferating and proliferating lymphoid cells. Nutr Cancer 38:131

    PubMed  Google Scholar 

  2. Borner C, Monney L (1999) Apoptosis without caspases: an inefficient molecular guillotine? Cell Death Differ 6:497

    Article  CAS  PubMed  Google Scholar 

  3. Bush JA, Cheung KJ Jr, Li G (2001) Curcumin induces apoptosis in human melanoma cells through a Fas receptor/caspase-8 pathway independent of p53. Exp Cell Res 271:305

    Article  CAS  PubMed  Google Scholar 

  4. Campone M, Vavasseur F, Le Cabellec MT, Meflah K, Vallette FM, Oliver L (2001) Induction of chemoresistance in HL-60 cells concomitantly causes a resistance to apoptosis and the synthesis of P-glycoprotein. Leukemia 15:1377

    Article  CAS  PubMed  Google Scholar 

  5. Cullen K, Davey R, Davey M (2001) The drug resistance proteins, multidrug resistance-associated protein and P-glycoprotein, do not confer resistance to Fas-induced cell death. Cytometry 43:189

    Article  CAS  PubMed  Google Scholar 

  6. Huang P, Oliff A (2001) Signaling pathways in apoptosis as potential targets for cancer therapy. Trends Cell Biol 11:343

    Article  CAS  PubMed  Google Scholar 

  7. Johnstone RW, Cretney E, Smyth MJ (1999) P-glycoprotein protects leukemia cells against caspase-dependent, but not caspase-independent, cell death. Blood 93:1075

    CAS  PubMed  Google Scholar 

  8. Johnstone RW, Ruefli AA, Tainton KM, Smyth MJ (2000) A role for P-glycoprotein in regulating cell death. Leuk Lymphoma 38:1

    CAS  PubMed  Google Scholar 

  9. Johnstone RW, Tainton KM, Ruefli AA, Froelich CJ, Cerruti L, Jane SM, Smyth MJ (2001) P-glycoprotein does not protect cells against cytolysis induced by pore- forming proteins. J Biol Chem 276:16667

    Article  CAS  PubMed  Google Scholar 

  10. Kim R, Tanabe K, Uchida Y, Emi M, Inoue H, Toge T (2002) Current status of the molecular mechanisms of anticancer drug-induced apoptosis. Cancer Chem Pharmacol 50:343

    Article  CAS  Google Scholar 

  11. Koizumi S, Konishi M, Ichihara T, Wada H, Matsukawa H, Goi K, Mizutani S (1995) Flow cytometric functional analysis of multidrug resistance by Fluo-3: a comparison with rhodamine-123. Eur J Cancer 31A:1682

    CAS  PubMed  Google Scholar 

  12. Krammer PH (2000) CD95’s deadly mission in the immune system. Nature 407:789

    Article  CAS  PubMed  Google Scholar 

  13. Kroemer G, Reed JC (2000) Mitochondrial control of cell death. Nat Med 6:513

    CAS  PubMed  Google Scholar 

  14. Lehne G (2000) P-glycoprotein as a drug target in the treatment of multidrug resistant cancer. Curr Drug Targets 1:85

    CAS  PubMed  Google Scholar 

  15. McGrath T, Center MS (1987) Adriamycin resistance in HL60 cells in the absence of detectable P-glycoprotein. Biochem Biophys Res Commun 145:1171

    CAS  PubMed  Google Scholar 

  16. McGrath T, Center MS (1988) Mechanisms of multidrug resistance in HL60 cells: evidence that a surface membrane protein distinct from P-glycoprotein contributes to reduced cellular accumulation of drug. Cancer Res 48:3959

    CAS  PubMed  Google Scholar 

  17. McGrath T, Latoud C, Arnold ST, Safa AR, Felsted RL, Center MS (1989) Mechanisms of multidrug resistance in HL60 cells. Analysis of resistance associated membrane proteins and levels of mdr gene expression. Biochem Pharmacol 38:3611

    CAS  PubMed  Google Scholar 

  18. Notarbartolo M, Cervello M, Dusonchet L, Cusimano A, D’Alessandro N (2002) Resistance to diverse apoptotic triggers in multidrug resistant HL60 cells and its possible relationship to the expression of P-glycoprotein, Fas and of the novel anti-apoptosis factors IAP (inhibitory of apoptosis proteins). Cancer Lett 180:91

    Article  CAS  PubMed  Google Scholar 

  19. Ogretmen B, Safa AR (1999) Negative regulation of MDR1 promoter activity in MCF-7, but not in multidrug resistant MCF-7/Adr, cells by cross-coupled NF-kappa B/p65 and c-Fos transcription factors and their interaction with the CAAT region. Biochemistry 38:2189

    Article  CAS  PubMed  Google Scholar 

  20. Piwocka K, Zablocki K, Wieckowski MR, Skierski J, Feiga I, Szopa J, Drela N, Wojtczak L, Sikora E (1999) A novel apoptosis-like pathway, independent of mitochondria and caspases, induced by curcumin in human lymphoblastoid T (Jurkat) cells. Exp Cell Res 249:299

    Article  CAS  PubMed  Google Scholar 

  21. Piwocka K, Bielak-Żmijewska A, Sikora E (2002) Curcumin induces caspase-3-independent apoptosis in human multidrug-resistant cells. Ann N Y Acad Sci 973:250

    CAS  PubMed  Google Scholar 

  22. Reed JC (2001) Apoptosis-regulating proteins as targets for drug discovery. Trends Mol Med 7:314

    Article  CAS  PubMed  Google Scholar 

  23. Ruefli AA, Smyth MJ, Johnstone RW (2000) HMBA induces activation of a caspase-independent cell death pathway to overcome P-glycoprotein-mediated multidrug resistance. Blood 95:2378

    PubMed  Google Scholar 

  24. Ruefli AA, Bernhard D, Tainton KM, Kofler R. Smyth M, Johnstone RW (2002) Suberoylanilide hydroxamic acid (SAHA) overcomes multidrug resistance and induces cell death in P-glycoprotein-expressing cells. Int J Cancer 99:292

    Article  CAS  PubMed  Google Scholar 

  25. Sikora E, Bielak-Zmijewska A, Piwocka K, Skierski J, Radziszewska E (1997) Inhibition of proliferation and apoptosis of human and rat T lymphocytes by curcumin, a curry pigment. Biochem Pharmacol 54:899

    Article  CAS  PubMed  Google Scholar 

  26. Woo JH, Kim YH, Choi YJ, Kim DG, Lee KS, Bae JH, Min DS, Chang JS, Jeong YJ, Lee YH, Park JW, Kwon TK (2003) Molecular mechanisms of curcumin-induced cytotoxicity: induction of apoptosis through generation of reactive oxygen species, down-regulation of Bcl-XL and IAP, the release of cytochrome c and inhibition of Akt. Carcinogenesis 24:1199

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by grants 6 P04A 03019 and 6 P04A 01519 from the Polish State Committee for Scientific Research (KBN), Poland.

Author information

Authors and Affiliations

  1. Laboratory of Molecular Bases of Aging, Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, 3 Pasteur St, 02-093, Warsaw, Poland

    Anna Bielak-Żmijewska, Katarzyna Piwocka, Adriana Magalska & Ewa Sikora

Authors
  1. Anna Bielak-Żmijewska
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Katarzyna Piwocka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Adriana Magalska
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ewa Sikora
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ewa Sikora.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bielak-Żmijewska, A., Piwocka, K., Magalska, A. et al. P-glycoprotein expression does not change the apoptotic pathway induced by curcumin in HL-60 cells. Cancer Chemother Pharmacol 53, 179–185 (2004). https://doi.org/10.1007/s00280-003-0705-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-003-0705-x

Keywords

Search

Navigation