Celecoxib inhibits MDR1 expression through COX-2-dependent mechanism in human hepatocellular carcinoma (HepG2) cell line
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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.
KeywordsAP-1 Celecoxib COX-2 HepG2 cells MDR1 Simulation
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.
- 1.Arino PA, Gottesman MM, Pastan I (1990) Regulation of multidrug resistance gene in regenerating rat liver. Cell Growth Differ 1:57–62Google 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 355:873–884CrossRefPubMedGoogle 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 52:237–245CrossRefPubMedGoogle 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 29:688–696CrossRefPubMedGoogle 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 48:69–79Google 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 121:605–617CrossRefPubMedGoogle 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 278:22199–22209CrossRefPubMedGoogle 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 43:433–442CrossRefPubMedGoogle Scholar