Phorbol Ester TPA Modulates Chemoresistance in the Drug Sensitive Breast Cancer Cell Line MCF-7 by Inducing Expression of Drug Efflux Transporter ABCG2

  • Kalalinia, Fatemeh (Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences) ;
  • Elahian, Fatemeh (Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences) ;
  • Hassani, Mitra (Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences) ;
  • Kasaeeian, Jamal (Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences) ;
  • Behravan, Javad (Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences)
  • Published : 2012.06.30


Recent studies have indicated a link between levels of cyclooxygenase-2 (COX-2) and development of the multidrug resistance (MDR) phenotype. The ATP-binding cassette sub-family G member 2 (ABCG2) is a major MDR-related transporter protein that is frequently overexpressed in cancer patients. In this study, we aimed to evaluate any positive correlation between COX-2 and ABCG2 gene expression using the COX-2 inducer 12-O-tetradecanoylphorbol-13-acetate (TPA) in human breast cancer cell lines. ABCG2 mRNA and protein expression was studied using real-time RT-PCR and flow cytometry, respectively. A significant increase of COX-2 mRNA expression (up to 11-fold by 4 h) was induced by TPA in MDA-MB-231 cells, this induction effect being lower in MCF-7 cells. TPA caused a considerable increase up to 9-fold in ABCG2 mRNA expression in parental MCF-7 cells, while it caused a small enhancement in ABCG2 expression up to 67 % by 4 h followed by a time-dependent decrease in ABCG2 mRNA expression in MDA-MB-231 cells. TPA treatment resulted in a slight increase of ABCG2 protein expression in MCF-7 cells, while a time-dependent decrease in ABCG2 protein expression was occurred in MDA-MB-231 cells. In conclusion, based on the observed effects of TPA in MDA-Mb-231 cells, it is proposed that TPA up-regulates ABCG2 expression in the drug sensitive MCF-7 breast cancer cell line through COX-2 unrelated pathways.


  1. Buchholz S, Keller G, Schally AV, et al (2006). Therapy of ovarian cancers with targeted cytotoxic analogs of bombesin, somatostatin, and luteinizing hormone-releasing hormone and their combinations. Proc Natl Acad Sci USA, 103, 10403-7.
  2. Cao Y, Prescott SM (2002). Many actions of cyclooxygenase-2 in cellular dynamics and in cancer. J Cell Physiol, 190, 279-86.
  3. Chan CC, Boyce S, Brideau C, et al (1995). Pharmacology of a selective cyclooxygenase-2 inhibitor, L-745,337: a novel nonsteroidal anti-inflammatory agent with an ulcerogenic sparing effect in rat and nonhuman primate stomach. J Pharmacol Exp Ther, 274, 1531-7.
  4. Chaudhary PM, Roninson IB (1992). Activation of MDR1 (P-glycoprotein) gene expression in human cells by protein kinase C agonists. Oncol Res, 4, 281-90.
  5. Diestra JE, Scheffer GL, Catala I, et al (2002). Frequent expression of the multi-drug resistance-associated protein BCRP/MXR/ABCP/ABCG2 in human tumours detected by the BXP-21 monoclonal antibody in paraffin-embedded material. J Pathol, 198, 213-9.
  6. Doyle LA, Ross DD (2003). Multidrug resistance mediated by the breast cancer resistance protein BCRP (ABCG2). Oncogene, 22, 7340-58.
  7. Drew L, Groome N, Warr JR, Rumsby MG (1996). Reduced daunomycin accumulation in drug-sensitive and multidrugresistant human carcinoma KB cells following phorbol ester treatment: a potential role for protein kinase C in reducing drug influx. Oncol Res, 8, 249-57.
  8. Dubois RN, Abramson SB, Crofford L, et al (1998). Cyclooxygenase in biology and disease. Faseb J, 12, 1063-73.
  9. Elahian F, Kalalinia F, Behravan J (2009). Dexamethasone downregulates BCRP mRNA and protein expression in breast cancer cell lines. Oncol Res, 18, 9-15.
  10. Elahian F, Kalalinia F, Behravan J (2010). Evaluation of indomethacin and dexamethasone effects on BCRPmediated drug resistance in MCF-7 parental and resistant cell lines. Drug Chem Toxicol, 33, 113-9.
  11. Fantappie O ME, Sardi I, Raimondi L, et al (2002 ). The MDR phenotype is associated with the expression of COX-2 and iNOS in a human hepatocellular carcinoma cell line. Hepatology, 35, 843-52.
  12. Fine RL, Patel J, Chabner BA (1988). Phorbol esters induce multidrug resistance in human breast cancer cells. Proc Natl Acad Sci USA, 85, 582-6.
  13. Huls M, Russel FG, Masereeuw R (2009). The role of ATP binding cassette transporters in tissue defense and organ regeneration. J Pharmacol Exp Ther, 328, 3-9.
  14. Kalalinia F, Elahian F, Behravan J (2010). Potential role of cyclooxygenase-2 on the regulation of the drug efflux transporter ABCG2 in breast cancer cell lines. J Cancer Res Clin Oncol, ?, ?-?.
  15. Kargman SL, O'Neill GP, Vickers PJ, et al (1995). Expression of prostaglandin G/H synthase-1 and -2 protein in human colon cancer. Cancer Res, 55, 2556-9.
  16. Lee JY, Tanabe S, Shimohira H, et al (2007). Expression of cyclooxygenase-2, P-glycoprotein and multi-drug resistanceassociated protein in canine transitional cell carcinoma. Res Vet Sci, 83, 210-6.
  17. Lee SH, Soyoola E, Chanmugam P, et al (1992). Selective expression of mitogen-inducible cyclooxygenase in macrophages stimulated with lipopolysaccharide. J Biol Chem, 267, 25934-8.
  18. Lin P, Hu SW, Chang TH (2003). Correlation between gene expression of aryl hydrocarbon receptor (AhR), hydrocarbon receptor nuclear translocator (Arnt), cytochromes P4501A1 (CYP1A1) and 1B1 (CYP1B1), and inducibility of CYP1A1 and CYP1B1 in human lymphocytes. Toxicol Sci, 71, 20-6.
  19. Liu XH, Rose DP (1996). Differential expression and regulation of cyclooxygenase-1 and -2 in two human breast cancer cell lines. Cancer Res, 56, 5125-7.
  20. Malekshah OM, Bahrami AR, Afshari JT, Mosaffa F, Behravan J (2011). Correlation Between PXR and ABCG2 Patterns of mRNA Expression in a MCF7 Breast Carcinoma Cell Derivative upon Induction by Proinflammatory Cytokines. DNA Cell Biol, 30, 25-31.
  21. Maliepaard M, Scheffer GL, Faneyte IF, et al (2001). Subcellular localization and distribution of the breast cancer resistance protein transporter in normal human tissues. Cancer Res, 61, 3458-64.
  22. Mao Q, Unadkat JD (2005). Role of the breast cancer resistance protein (ABCG2) in drug transport. Aaps J, 7, 118-33.
  23. Minderman H, Suvannasankha A, O'Loughlin KL, et al (2002). Flow cytometric analysis of breast cancer resistance protein expression and function. Cytometry, 48, 59-65.
  24. Mosaffa F, Lage H, Afshari JT, Behravan J (2009). Interleukin-1 beta and tumor necrosis factor-alpha increase ABCG2 expression in MCF-7 breast carcinoma cell line and its mitoxantrone-resistant derivative, MCF-7/MX. Inflamm Res, 58, 669-76.
  25. O'Neill GP, Ford-Hutchinson AW (1993). Expression of mRNA for cyclooxygenase-1 and cyclooxygenase-2 in human tissues. FEBS Lett, 330, 156-60.
  26. Patel VA, Dunn MJ, Sorokin A (2002). Regulation of MDR1 (P-glycoprotein) by cyclooxygenase-2. J Biol Chem, 277, 38915-20.
  27. Puhlmann U, Ziemann C, Ruedell G, et al (2005). Impact of the cyclooxygenase system on doxorubicin-induced functional multidrug resistance 1 overexpression and doxorubicin sensitivity in acute myeloid leukemic HL-60 cells. J Pharmacol Exp Ther, 312, 346-54.
  28. Ramachandran C, Kunikane H, You W, Krishan A (1998). Phorbol ester-induced P-glycoprotein phosphorylation and functionality in the HTB-123 human breast cancer cell line. Biochem Pharmacol, 56, 709-18.
  29. Rapisarda A, Uranchimeg B, Scudiero DA, et al (2002). Identification of small molecule inhibitors of hypoxiainducible factor 1 transcriptional activation pathway. Cancer Res, 62, 4316-24.
  30. Ratnasinghe D, Daschner PJ, Anver MR, et al (2001). Cyclooxygenase-2, P-glycoprotein-170 and drug resistance; is chemoprevention against multidrug resistance possible? Anticancer Res, 21, 2141-7.
  31. Ross DD, Karp JE, Chen TT, Doyle LA (2000). Expression of breast cancer resistance protein in blast cells from patients with acute leukemia. Blood, 96, 365-8.
  32. Shen SC, Ko CH, Hsu KC, Chen YC (2004). 3-OH flavone inhibition of epidermal growth factor-induced proliferaton through blocking prostaglandin E2 production. Int J Cancer, 108, 502-10.
  33. Smith WL, Dewitt DL (1996). Prostaglandin endoperoxide H synthases-1 and -2. Adv Immunol, 62, 167-215.
  34. Sorokin A (2004). Cyclooxygenase-2: potential role in regulation of drug efflux and multidrug resistance phenotype. Curr Pharm Des, 10, 647-57.
  35. Surowiak P, Materna V, Denkert C, et al (2006). Significance of cyclooxygenase 2 and MDR1/P-glycoprotein coexpression in ovarian cancers. Cancer Lett, 235, 272-80.
  36. Surowiak P, Materna V, Matkowski R, et al (2005). Relationship between the expression of cyclooxygenase 2 and MDR1/Pglycoprotein in invasive breast cancers and their prognostic significance. Breast Cancer Res, 7, 862-70.
  37. Surowiak P, Pawelczyk K, Maciejczyk A, et al (2008). Positive correlation between cyclooxygenase 2 and the expression of ABC transporters in non-small cell lung cancer. Anticancer Res, 28, 2967-74.
  38. Tanioka T, Nakatani Y, Kobayashi T, et al (2003). Regulation of cytosolic prostaglandin E2 synthase by 90-kDa heat shock protein. Biochem Biophys Res Commun, 303, 1018-23.
  39. Trebino CE, Stock JL, Gibbons CP, et al (2003). Impaired inflammatory and pain responses in mice lacking an inducible prostaglandin E synthase. Proc Natl Acad Sci U S A, 100, 9044-9.

Cited by

  1. ABCG2 gene amplification and expression in esophageal cancer cells with acquired adriamycin resistance vol.9, pp.4, 2014,
  2. ABCG2 inhibition as a therapeutic approach for overcoming multidrug resistance in cancer vol.41, pp.2, 2016,
  3. The multiple facets of drug resistance: one history, different approaches vol.33, pp.1, 2014,