Skip to main content

Advertisement

SpringerLink
Log in

Effect of MDR modulators verapamil and promethazine on gene expression levels of MDR1 and MRP1 in doxorubicin-resistant MCF-7 cells

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

Abstract

Purpose

One of the major problems of cancer chemotherapy is the development of multidrug resistance (MDR) phenotype. Among the numerous mechanisms of MDR, a prominent one is the increased expression of membrane transporter proteins, the action of which leads to decreased intracellular drug concentration and cytotoxicity of drugs. Among them, P-gp and MRP1, encoded by MDR1 and MRP1 genes, respectively, have been associated with MDR phenotype. Chemical modulators can be used to reverse MDR. These chemicals can either modulate MDR due to their substrate analogy (such as calcium channel blocker verapamil) or interact with phospholipid membranes (such as antihistaminic drug promethazine). This study focuses on the effect of verapamil and promethazine on the expression levels of MDR1 and MRP1 genes and the drug transport activity in doxorubicin-resistant MCF-7 breast carcinoma cell line.

Methods

Doxorubicin-resistant MCF-7 (MCF-7/Dox) cells were incubated with either verapamil or promethazine, and total RNA was isolated. Real-time PCR (qPCR) was carried out by using specific primers for MDR1, MRP1, and ß-actin genes. Intracellular doxorubicin accumulation was also examined by confocal laser scanning microscopy in treated cells.

Results

Results demonstrated a significant decrease in both MDR1 and MRP1 expression levels after promethazine applications. It has also been shown that treatment of the cells with verapamil results in significant decrease in MDR1 mRNA levels. Confocal laser scanning microscopy images demonstrated that the intracellular accumulation of doxorubicin was increased after verapamil treatment in MCF-7/Dox cells.

Conclusions

The present study gives an idea about the efficiency of verapamil and promethazine on MDR reversal both in gene expression and in transport activity levels.

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

Similar content being viewed by others

References

  1. Ambudkar S, Kimchi-Sarfaty C, Sauna Z, Gottesman M (2003) P-glycoprotein: from genomics to mechanism. Oncogene 22:7468–7485

    Article  PubMed  CAS  Google Scholar 

  2. Anuchapreeda S, Leechanachai P, Smith MM, Ambudkar SV, Limtrakul PN (2002) Modulation of P-glycoprotein expression and function by curcumin in multidrug-resistant human KB cells. Biochem Pharmacol 64:573–582

    Article  PubMed  CAS  Google Scholar 

  3. Callaghan R, Stafford A, Epand RM (1993) Increased accumulation of drugs in a multidrug resistant cell line by alteration of membrane biophysical properties. Biochim Biophys Acta 1175:277–282

    Article  PubMed  CAS  Google Scholar 

  4. Dalton WS (1997) Mechanisms of drug resistance in hematologic malignancies. Semin Hematol 34:3–8

    PubMed  CAS  Google Scholar 

  5. Drori S, Eytan GD, Assaraf YG (1995) Potentiation of anticancer drug cytotoxicity by multidrug resistance chemosensitizers involves alterations in membrane fluidity leading to increased membrane permeability. Eur J Biochem 228:1020–1029

    Article  PubMed  CAS  Google Scholar 

  6. Eskiocak U, İşeri ÖD, Kars MD, Biçer A, Gündüz U (2008) Effect of doxorubicin on telomerase activity and apoptotic gene expression in doxorubicin resistant and sensitive MCF-7 cells. Chemotherapy 54:209–216

    Article  PubMed  CAS  Google Scholar 

  7. Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2(1):48–58

    Article  PubMed  CAS  Google Scholar 

  8. Hilgeroth A, Molnár A, Molnár J, Voigt B (2006) Correlation of calculated molecular orbital energies of some phenothiazine compounds with MDR reversal properties. Eur J Med Chem 41:548–551

    Article  PubMed  CAS  Google Scholar 

  9. İşeri ÖD, Kars MD, Eroğlu S, Gündüz U (2009) Cross-resistance development and combined applications of anticancer agents in drug resistant MCF-7 cell lines. Int J Hem Oncol 1(19):1–8

    Google Scholar 

  10. Kars MD, Işeri OD, Gündüz U, Ural AU, Arpaci F, Molnar J (2006) Development of rational in vitro models for drug resistance in breast cancer and modulation of MDR by selected compounds. Anticancer Res 26:4559–4568

    PubMed  CAS  Google Scholar 

  11. Kars MD, Iseri OD, Gunduz U, Molnar J (2008) Reversal of MDR by synthetic and natural compounds in drug resistant MCF-7 cell lines. Chemotherapy 54:194–200

    Article  PubMed  CAS  Google Scholar 

  12. Kars MD, Iseri ÖD, Ural AU, Avcu F, Beyzadeoglu M, Dirican B, Gündüz U (2009) Development of radioresistance in drug resistant human MCF-7 breast cancer cells. J Radiol Pract. doi:10.1017/S1460396909990070

  13. Kruh GD, Belinsky MG (2003) The MRP family of drug efflux pumps. Oncogene 22:7537–7552

    Article  PubMed  CAS  Google Scholar 

  14. Litman T, Druley TE, Stein WD, Bates SE (2001) From MDR to MXR: new understanding of multidrugresistance systems, their properties and clinical significance. CMLS Cell Mol Life Sci 58:931–959

    Article  CAS  Google Scholar 

  15. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and 2−∆∆CT method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  16. Michalak K, Wesołowska O, Motohashi N, Hendrich AB (2007) The role of the membrane actions of phenothiazines and flavonoids as functional modulators. Top Heterocycl Chem 8:223–302

    Article  CAS  Google Scholar 

  17. Molnar J, Gyémánt N, Tanaka M, Hohmann J, Bergmann-Leitner E, Molnár P, Deli J, Didiziapetris R, Ferreira M-JU (2006) Inhibition of multidrug resistance of cancer cells by natural diterpens, triterpenes and carotenoids. Curr Pharm Design 12:287–311

    Article  CAS  Google Scholar 

  18. Molnar J, Szabo D, Mandi Y, Mucsi I, Fischer J, Varga A, Konig S, Motohashi N (1998) Multidrug resistance reversal in mouse lymphoma cells by heterocyclic compounds. Anticancer Res 18:3033–3038

    PubMed  CAS  Google Scholar 

  19. Motohashi N, Kurihara T, Wakabayashi H, Yaji M, Mucsi I, Molnar J, Maruyama S, Sakagami H, Nakashima H, Tani S, Shirataki Y, Kawase M (2001) Biological activity of a fruit vegetable, “Anastasia green”, a species of sweet pepper. In Vivo 5:437–442

    Google Scholar 

  20. Muller C, Goubin F, Ferrandis E, Cornil-Scharwtz I, Bailly JD, Bordier C, Bénard J, Sikic BI, Laurent G (1995) Evidence for transcriptional control of human MDR1 gene expression by verapamil in multidrug-resistant leukemic cells. Mol Pharmacol 47(1):51–56

    PubMed  CAS  Google Scholar 

  21. Mutoh K, Tsukahara S, Mitsuhashi J, Katayama K, Sugimoto Y (2006) Estrogen-mediated post transcriptional down-regulation of P-glycoprotein in MDR1-transduced human breast cancer cells. Cancer Sci 97:1198–1204

    Article  PubMed  CAS  Google Scholar 

  22. Nobili S, Landini I, Giglioni B, Mini E (2006) Pharmacological strategies for overcoming multidrug resistance. Curr Drug Targets 7(7):861–879

    Article  PubMed  CAS  Google Scholar 

  23. Pajeva I, Wiese M (1998) Molecular modeling of phenothiazines and related drugs as multidrug resistance modifiers: a comparative molecular field analysis study. J Med Chem 41:1815–1826

    Article  PubMed  CAS  Google Scholar 

  24. Perez E (2009) Impact, mechanisms, and novel chemotherapy strategies for overcoming resistance to anthracyclines and taxanes in metastatic breast cancer. Breast Can Res Treat 114:195–201

    Article  CAS  Google Scholar 

  25. Schuldes H, Dolderer J, Knobloch L, Bade S, Bickeboeller R, Woodcock BG, Jonas D, Zimmer G (1998) Relationship between plasma membrane fluidity and R-verapamil action in CHO cells. Int J Clin Pharmacol Ther 36:71–73

    PubMed  CAS  Google Scholar 

  26. Trock B, Leonessa F, Clarke R (1997) Multidrug resistance in breast cancer: a meta-analysis of MDR1/gp170 expression and its possible functional significance. J Nat Can Inst 89:917–931

    Article  CAS  Google Scholar 

  27. Young AM, Allen CE, Audus KL (2003) Efflux transporters of the human placenta. Adv Drug Deliver Rev 55:125–132

    Article  CAS  Google Scholar 

  28. Yu ST, Chen TM, Tseng SY, Chen YH (2007) Tryptanthrin inhibits MDR1 and reverses doxorubicin resistance in breast cancer cells. Biochem Biophys Res Commun 358:79–84

    Article  PubMed  CAS  Google Scholar 

  29. Zhou YG, Li KY, Li HD (2008) Effect of the novel antipsychotic drug perospirone on P-glycoprotein function and expression in Caco-2 cells. Eur J Clin Pharmacol 64(7):697–703

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Dr. Can Özen (Middle East Technical University Central Laboratory) is greatly acknowledged for his contributions in laser scanning microscopy and image analysis.

Author information

Authors and Affiliations

  1. Department of Biological Sciences, Middle East Technical University, 06531, Ankara, Turkey

    Yaprak Dönmez, Laila Akhmetova, Meltem Demirel Kars & Ufuk Gündüz

  2. Institute of Transplantation and Gene Sciences, Başkent University, 06980, Ankara, Turkey

    Özlem Darcansoy İşeri

Authors
  1. Yaprak Dönmez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laila Akhmetova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Özlem Darcansoy İşeri
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Meltem Demirel Kars
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ufuk Gündüz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ufuk Gündüz.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dönmez, Y., Akhmetova, L., İşeri, Ö.D. et al. Effect of MDR modulators verapamil and promethazine on gene expression levels of MDR1 and MRP1 in doxorubicin-resistant MCF-7 cells. Cancer Chemother Pharmacol 67, 823–828 (2011). https://doi.org/10.1007/s00280-010-1385-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-010-1385-y

Keywords

Search

Navigation