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Celecoxib inhibits MDR1 expression through COX-2-dependent mechanism in human hepatocellular carcinoma (HepG2) cell line

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Abstract

The role of COX-2 in the regulation of the expression of MDR1, a P-glycoprotein involved in hepatocellular carcinoma cell line, HepG2, was studied in the present investigation. Celecoxib, a selective inhibitor of COX-2, at 25 μM concentration increased the accumulation of doxorubicin in HepG2 cells and enhanced the sensitivity of the cells to doxorubicin by tenfold. The induction of MDR1 expression by PGE2 and its downregulation by celecoxib or by COX-2 knockdown suggests that the enhanced sensitivity of HepG2 cells to doxorubicin by celecoxib is mediated by the downregulation of MDR1 expression, through COX-2-dependent mechanism. Further studies revealed the involvement of AP-1 in the celecoxib-induced downregulation of MDR1 expression. These experimental studies correlated well with in silico predictions and further suggested the inactivation of the signal transduction pathways involving ERK, JNK and p38. The present study thus demonstrates the usefulness of COX-2 intervention in overcoming the drug resistance in HepG2 cells.

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References

  1. Arino PA, Gottesman MM, Pastan I (1990) Regulation of multidrug resistance gene in regenerating rat liver. Cell Growth Differ 1:57–62

    Google Scholar 

  2. Bertagnolli MM, Eagle CJ, Zauber AG, Redston M, Solomon SD, Kim K, Tang J, Rosenstein RB, Wittes J, Corle D, Hess TM, Woloj GM, Boisserie F, Anderson WF, Viner JL, Bagheri D, Burn J, Chung DC, Dewar T, Foley TR, Hoffman N, Macrae F, Pruitt RE, Saltzman JR, Salzberg B, Sylwestrowicz T, Gordon GB, Hawk ET (2006) Celecoxib for the prevention of sporadic colorectal adenomas. N Engl J Med 355:873–884

    Article  PubMed  CAS  Google Scholar 

  3. Bhushan A, Abramson R, Chiu JF, Tritton TR (1992) Expression of c-fos in human and murine multidrug-resistant cells. Mol Pharmacol 42:69–74

    PubMed  CAS  Google Scholar 

  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  5. Bradley G, Sharma R, Rajalakshmi S, Ling V (1992) P-glycoprotein expression during tumor progression in the rat liver. Cancer Res 52:5154–5161

    PubMed  CAS  Google Scholar 

  6. Campling BG, Pym J, Galbraith PR, Cole SP (1988) Use of the MTT assay for rapid determination of chemosensitivity of human leukemic blast cells. Leuk Res 12:823–831

    Article  PubMed  CAS  Google Scholar 

  7. Chan JY, Chu AC, Fung KP (2000) Inhibition of P-glycoprotein expression and reversal of drug resistance of human hepatoma HepG2 cells by multidrug resistance gene (mdr1) antisense RNA. Life Sci 67:2117–2124

    Article  PubMed  CAS  Google Scholar 

  8. Chen Y, Hughes-Fulford M (2000) Prostaglandin E2 and the protein kinase A pathway mediate arachidonic acid induction of c-fos in human prostate cancer cells. Br J Cancer 82:2000–2006

    Article  PubMed  CAS  Google Scholar 

  9. Dalle S, Longuet C, Costes S, Broca C, Faruque O, Fontes G (2004) Glucagon promotes cAMP-response element-binding protein phosphorylation via activation of ERK1/2 in MIN6 cell line and isolated islets of Langerhans. J Biol Chem 279:20345–20355

    Article  PubMed  CAS  Google Scholar 

  10. Daschner PJ, Ciolino HP, Plouzek CA, Yeh GC (1999) Increased AP-1 activity in drug resistant human breast cancer MCF-7 cells. Breast Cancer Res Treat 53:229–240

    Article  PubMed  CAS  Google Scholar 

  11. Eberhart CE, Coffey RJ, Radhika A, Giardiello FM, Ferrenbach S, DuBois RN (1994) Up-regulation of cyclooxygenase 2 gene expression in human colorectal adenomas and adenocarcinomas. Gastroenterology 107:1183–1188

    PubMed  CAS  Google Scholar 

  12. Fujino H, Salvi S, Regan JW (2005) Differential regulation of phosphorylation of the cAMP response element-binding protein after activation of EP2 and EP4 prostanoid receptors by prostaglandin E2. Mol Pharmacol 68:251–259

    PubMed  CAS  Google Scholar 

  13. Gallagher ED, Xu S, Moomaw C, Slaughter CA, Cobb MH (2002) Binding of JNK/SAPK to MEKK1 is regulated by phosphorylation. J Biol Chem 277:45785–45792

    Article  PubMed  CAS  Google Scholar 

  14. Hairer E, Wanner G (1999) Stiff differential solved by Radau methods. J Comput Appl Math 111:93–111

    Article  Google Scholar 

  15. Hanif R, Pittas A, Feng Y, Koutsos MI, Qiao L, Staiano-Coico L, Shiff SI, Rigas B (1996) Effects of nonsteroidal anti-inflammatory drugs on proliferation and on induction of apoptosis in colon cancer cells by a prostaglandin-independent pathway. Biochem Pharmacol 52:237–245

    Article  PubMed  CAS  Google Scholar 

  16. Huang C, Wu M, Xu G, Li D, Cheng H, Tu ZX, Jiang HQ, Gu JR (1992) Over expression of the mdr1 gene and P-glycoprotein in human hepatocellular carcinoma. J Natl Cancer Inst 84:262–264

    Article  PubMed  CAS  Google Scholar 

  17. Johnstone RW, Ruefli AA, Tainton KM, Smyth MJ (2000) A role for P-glycoprotein in regulating cell death. Leuk Lymphoma 38:1–11

    PubMed  CAS  Google Scholar 

  18. Kataoka K, Takikawa Y, Lin SD, Suzuki K (2005) Prostaglandin E2 receptor EP4 agonist induces Bcl-xL and independently activates proliferation signals in mouse primary hepatocytes. J Gastroenterol 40:610–616

    Article  PubMed  CAS  Google Scholar 

  19. Kawamori T, Rao CV, Seibert K, Reddy BS (1998) Chemopreventive activity of celecoxib, a specific cyclooxygenase-2 inhibitor, against colon carcinogenesis. Cancer Res 58:409–412

    PubMed  CAS  Google Scholar 

  20. Koga H, Sakisaka S, Ohishi M, Kawaguchi T, Taniguchi E, Sasatomi K, Harada M, Kusaba T, Tanaka M, Kimura R, Nakashima Y, Nakashima O, Kojiro M, Kurohiji T, Sata M (1999) Expression of cyclooxygenase-2 in human hepatocellular carcinoma: relevance to tumor differentiation. Hepatology 29:688–696

    Article  PubMed  CAS  Google Scholar 

  21. Kotani M, Tanaka I, Ogawa Y, Usui T, Mori K, Ichikawa A, Narumiya S, Yoshimi T, Nakao K (1995) Molecular cloning and expression of multiple isoforms of human prostaglandin E receptor EP3 subtype generated by alternative messenger RNA splicing: multiple second messenger systems and tissue-specific distributions. Mol Pharmacol 48:69–79

    Google Scholar 

  22. Marroni M, Agrawal ML, Kight K, Hallene KL, Hossain M, Cucullo L, Signorelli K, Namura S, Bingaman W, Janigro D (2003) Relationship between expression of multiple drug resistance proteins and p53 tumor suppressor gene proteins in human brain astrocytes. Neuroscience 121:605–617

    Article  PubMed  CAS  Google Scholar 

  23. Nagasue N, Dhar DK, Makino Y, Yoshimura H, Nakamura T (1995) Over expression of P-glycoprotein in adenomatous hyperplasia of human liver with cirrhosis. J Hepatol 22:197–201

    Article  PubMed  CAS  Google Scholar 

  24. Ouellet M, Falgueyret JP, Ear PH, Pen A, Mancini JA, Riendeau D (2002) Purification and characterization of recombinant microsomal prostaglandin E synthase-1. Protein Expr Purif 26:489–495

    Article  PubMed  CAS  Google Scholar 

  25. Park JG, Lee SK, Hong IG, Kim HS, Lim KH, Choe KJ, Kim WH, Kim YI, Tsuruo T, Gottesman MM (1994) MDR1 gene expression: its effect on drug resistance to doxorubicin in human hepatocellular carcinoma cell lines. J Natl Cancer Inst 86:700–705

    Article  PubMed  CAS  Google Scholar 

  26. Patel VA, Dunn MJ, Sorokin A (2002) Regulation of MDR-1 (P-glycoprotein) by cyclooxygenase-2. J Biol Chem 277:38915–38920

    Article  PubMed  CAS  Google Scholar 

  27. Roesler WJ, Simard J, Graham JG, McFie PJ (1994) Characterization of the liver-specific component of the cAMP response unit in the phosphoenolpyruvate carboxykinase (GTP) gene promoter. J Biol Chem 269:14276–14283

    PubMed  CAS  Google Scholar 

  28. Roller A, Bahr OR, Streffer J, Winter S, Heneka M, Deininger M, Meyermann R, Naumann U, Gulbins E, Weller M (1999) Selective potentiation of drug cytotoxicity by NSAID in human glioma cells: the role of COX-1 and MRP. Biochem Biophys Res Commun 259:600–605

    Article  PubMed  CAS  Google Scholar 

  29. Schrenk D, Gant TW, Preisegger KH, Silverman JA, Marino PA, Thorgeirsson SS (1993) Induction of multidrug resistance gene expression during cholestasis in rats and non human primates. Hepatology 17:854–860

    Article  PubMed  CAS  Google Scholar 

  30. Shaik MS, Chatterjee A, Jackson T, Singh M (2006) Enhancement of anti-tumor activity of docetaxel by celecoxib in lung tumors. Int J Cancer 118:396–404

    Article  PubMed  CAS  Google Scholar 

  31. Silverman JA, Schrenk D (1997) Expression of the multidrug resistance genes in the liver. FASEB J 11:308–313

    PubMed  CAS  Google Scholar 

  32. Silverman JA, Thorgeirsson SS (1995) Regulation and function of the multidrug resistance genes in liver. Prog Liver Dis 13:101–123

    PubMed  CAS  Google Scholar 

  33. Sukhai M, Piquette-Miller M (2000) Regulation of the multidrug resistance genes by stress signals. J Pharm Pharm Sci 3:268–280

    PubMed  CAS  Google Scholar 

  34. Thoren S, Weinander R, Saha S, Jegerschold C, Pettersson PL, Samuelsson B, Hebert H, Hamberg M, Morgenstern R, Jakobsson PJ (2003) Human microsomal prostaglandin E synthase-1: purification, functional characterization, and projection structure determination. J Biol Chem 278:22199–22209

    Article  PubMed  CAS  Google Scholar 

  35. Tucker ON, Dannenberg AJ, Yang EK, Zhang F, Teng L, Daly JM, Soslow R, Masferrer JL, Woerner BM, Koki AT, Fahey TJ III (1999) Cyclooxygenase-2 expression is up-regulated in human pancreatic cancer. Cancer Res 59:987–990

    PubMed  CAS  Google Scholar 

  36. van Wijngaarden J, van Beek E, van Rossum G, van der Bent C, Hoekman K, van der Pluijm G, van der Pol MA, Broxterman HJ, van Hinsbergh VW, Lowik CW (2007) Celecoxib enhances doxorubicin-induced cytotoxicity in MDA-MB231 cells by NF-kappaB-mediated increase of intracellular doxorubicin accumulation. Eur J Cancer 43:433–442

    Article  PubMed  CAS  Google Scholar 

  37. Vogel C (2000) Prostaglandin H synthases and their importance in chemical toxicity. Curr Drug Metab 1:391–404

    Article  PubMed  CAS  Google Scholar 

  38. Wolff H, Saukkonen K, Anttila S, Karjalainen A, Vainio H, Ristimaki A (1998) Expression of cyclooxygenase-2 in human lung carcinoma. Cancer Res 58:4997–5001

    PubMed  CAS  Google Scholar 

  39. Xu L, Han C, Lim K, Wu T (2006) Cross-talk between peroxisome proliferator-activated receptor delta and cytosolic phospholipase A(2)alpha/cyclooxygenase-2/prostaglandin E(2) signaling pathways in human hepatocellular carcinoma cells. Cancer Res 66:11859–11868

    Article  PubMed  CAS  Google Scholar 

  40. Ziemann C, Schafer D, Rudell G, Kahl GF, Hirsch-Ernst KI (2002) The cyclooxygenase system participates in functional mdr1b overexpression in primary rat hepatocyte cultures. Hepatology 35:579–588

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by research grants from Council of Scientific and Industrial Research (CSIR) (Grant # 37(1221)/05/EMR-II) and from Department of Science and Technology (DST) (Grant # VII-PRDSF/50/05-06/TDT), Govt. of India. We duly acknowledge CSIR, Govt. of India for providing Senior Research Fellowship to Dr. Karnati R. Roy and Dr. Smita Agarwal; University Grants Commission (UGC), New Delhi for providing Dr. D. S. Kothari-Postdoctoral Fellowship (PDF) to Dr. Gorla V. Reddy; DBT, Govt. of India for providing UOH-CREBB-Junior Research Fellowship (JRF) to Chandrani Achari. We thank Mr. Ch. Shiva Kumar, technical assistant, for his help during the manuscript preparation.

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Authors and Affiliations

  1. Department of Animal Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, 500046, India

    Karnati R. Roy, Gorla V. Reddy, Smita Agarwal, Chandrani Achari & Pallu Reddanna

  2. Cellworks Group Inc., California, USA

    Leela Maitreyi & Shireen Vali

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  1. Karnati R. Roy
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Correspondence to Pallu Reddanna.

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K. R. Roy and G. V. Reddy contributed equally to this work.

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Roy, K.R., Reddy, G.V., Maitreyi, L. et al. Celecoxib inhibits MDR1 expression through COX-2-dependent mechanism in human hepatocellular carcinoma (HepG2) cell line. Cancer Chemother Pharmacol 65, 903–911 (2010). https://doi.org/10.1007/s00280-009-1097-3

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