, Volume 27, Issue 1, pp 89–98 | Cite as

Antineoplastic influence of nimesulide in chemically induced hepatocellular carcinoma by inhibition of DNA synthesis

  • M. Afzal
  • D. P. Bhardwaj
  • R. Khan
  • Imran KazmiEmail author
  • S. Saleem
  • F. A. Al-Abbasi
  • Firoz AnwarEmail author
Original Article


Hepatocellular carcinoma is emerging as one of the most common forms of cancer resulting in thousands of death worldwide. The purpose of this study was to screen nimesulide for anticancer activity in chemically induced hepatocellular carcinoma in Wistar rats as well as in BEL 7402 and HEP G2 cell lines. HCC in rats was induced by administering a single dose of diethyl nitrosamine (150 mg/kg) intraperitoneally. Duration of the in vivo study was 12 weeks and the anticancer potential was further confirmed by in vitro cell line study. Administration of DENA in Wistar rats significantly elevated the levels of serum biochemical parameters and α-feto protein. Treatment with different dose of nimesulide significantly decreased the markedly raised serum levels of biochemical parameters as well as maintained the histology of the liver tissues nearly similar to the normal. Further study of hepatocytes enzymes showed that treatment with nimesulide also improved the antioxidant enzyme levels. Our study also examined the cytotoxicity and DNA synthesis inhibition by nimesulide in BEL 7402 and Hep G2 cell lines. Cell viability was assessed by [3H]-thymidine uptake procedure. The results obtained by in vitro cell line study, histopathological and biochemical data concluded that nimesulide, a preferential COX-2 inhibitor, has anticancer activity, which is by first reducing the formation of reactive oxygen species and second by inhibiting the PGE2 effect via Wnt signaling pathway (cell invasion, angiogenesis, and cell proliferation).


COX-2 inhibitor Diethylnitrosamine Biochemical parameters Hepatoprotective 



Hepatocellular carcinoma


Alkaline phosphatase


Total bilirubin


α-Feto protein



This research work was not funded by any organization.

Compliance with ethical standards

Conflict of interest

The author declares that there is no competing interest.

Compliance with ethics requirements

Manuscript is under compliance with ethical standard of journal.


  1. Abel S, DeKock M, van Schalkwyk DJ, Swanevelder S, Kew MC, Gelderblom WC (2009) Altered lipid profile, oxidative status and hepatitis B virus interactions in human hepatocellular carcinoma. Prost Leukot Essent Fatty Acids 81:391–399CrossRefGoogle Scholar
  2. Afzal M, Kazmi I, Gupta G, Rahman M, Kimothi V, Anwar F (2012) Preventive effect of metformin against N-nitrosodiethylamine-initiated hepatocellular carcinoma in rats. Saudi Pharm J 20:365–370CrossRefGoogle Scholar
  3. Al-Rejaie SS, Aleisa AM, Al-Yahya AA, Bakheet SA, Alsheikh A, Fatani AG, Al-Shabanah OA, Sayed-Ahmed MM (2009) Progression of diethylnitrosamine-induced hepatic carcinogenesis in carnitine-depleted rats. World J Gastroenterol 151:373–380Google Scholar
  4. Alwahaibi N, Mohamed J, Alhamadani A (2010) Supplementation of selenium reduces chemical hepatocarcinogenesis in male Sprague-Dawley rats. J Trace Elem Med Biol 24:119–123CrossRefGoogle Scholar
  5. Bansal AK, Bansal M, Soni G, Bhatnagar D (2005) Protective role of Vitamin E pre-treatment on N-nitrosodiethylamine induced oxidative stress in rat liver. Chem Biol Interact 156:101–111CrossRefGoogle Scholar
  6. Barbisan LF, Scolastici C, Miyamoto M, Salvadori DM, Ribeiro LR, da Eira AF, de Camargo JL (2003) Effects of crude extracts of Agaricus blazei on DNA damage and on rat liver carcinogenesis induced by diethylnitrosamine. Gen Mol Res 2:295–308Google Scholar
  7. Boland GP, Butt IS, Prasad R, Knox WF, Bundred NJ (2004) COX-2 expression is associated with an aggressive phenotype in ductal carcinoma in situ. Br J Cancer 90:423–429CrossRefGoogle Scholar
  8. Buckman SY, Gresham A, Hale P (1998) COX-2 expression is induced by UVB exposure in human skin: implications for the development of skin cancer. Carcinogenesis 19:723–729CrossRefGoogle Scholar
  9. Ding WQ, Liu B, Vaught JL, Yamauchi H, Lind SE (2005) Anticancer activity of the antibiotic clioquinol. Cancer Res 65:3389–3395CrossRefGoogle Scholar
  10. El-Serag HB, Mason AC (1999) Rising incidence of hepatocellular carcinoma in the United States. N Engl J Med 340:745–750CrossRefGoogle Scholar
  11. Esteves-Souza K, Pissinate MG, Nascimento NF, Grynberg A, Echevarria A (2006) Synthesis, cytotoxicity, and DNA-topoisomerase activity of new asymmetric ureas and thioureas. J Med Chem 14:492–499Google Scholar
  12. Fodera DD, Alessandro N, Cusimano A, Poma P, Notarbartolo M, Lampiasi N, Montalto G, Cervello M (2004) Induction of apoptosis and inhibition of cell growth in human hepatocellular carcinoma cells by COX-2 inhibitors. Ann N Y Acad 1028:440–449CrossRefGoogle Scholar
  13. Ghorab MM, Ragab FA, Hamed MM (2009) Design, synthesis and anticancer evaluation of novel tetrahydroquinoline derivatives containing sulfonamide moiety. Eur J Med Chem 44:4211–4217CrossRefGoogle Scholar
  14. Ghouri YA, Mian I, Rowe JH (2017) Review of hepatocellular carcinoma: epidemiology, etiology, and carcinogenesis. J Carcinog 16:1CrossRefGoogle Scholar
  15. Hida T, Kozaki K, Muramatsu H, Masuda A, Shimizu S, Mitsudomi T, Sugiura T, Ogawa M, Takahashi T (2000) Cyclooxygenase-2 inhibitor induces apoptosis and enhances cytotoxicity of various anticancer agents in non-small cell lung cancer cell lines. Clin Cancer Res 6:2006–2011Google Scholar
  16. Jiang J, Nilsson-Ehle P, Xu N (2006) Influence of liver cancer on lipid and lipoprotein metabolism. Lipids Health Dis 5:4CrossRefGoogle Scholar
  17. Kaplan MM (1993) Laboratory tests. In: Schiff L, Schiff ER (eds) Diseases of the liver, 7th edn. J B Lippincott, Philadelphia, pp 108–144Google Scholar
  18. Keller JJ, Giardiello FM (2003) Chemoprevention strategies using NSAIDs and COX- 2 inhibitors. Cancer Biol Ther 2:140–149CrossRefGoogle Scholar
  19. Koki AT, Masferre JL (2002) Celecoxib: a specific cox-2 inhibitor with anticancer properties. Cancer Control 9:28–35CrossRefGoogle Scholar
  20. Liang M, Yang H, Fu J (2009) Nimesulide inhibits IFN-c-induced programmed death-1-ligand 1surface expression in breast cancer cells by COX-2 and PGE2 independent mechanisms. Cancer Lett 276:47–52CrossRefGoogle Scholar
  21. Muzio G, Marengo B, Salvo R, Semeraro A, Canuto RA, Tessitore L (1999) Liver cancer is induced by a subnecrogenic dose of DENA when associated with fasting/refeeding: role of glutathione-transferase and lipid peroxidation. Free Rad Biol Med 26:1314–1320CrossRefGoogle Scholar
  22. Parkin DM, Bray F, Ferlay J (2001) Estimating the world cancer burden: globocan 2000. Int J Cancer 94:153–156CrossRefGoogle Scholar
  23. Plaa GL, Hewitt WR (1989) Detection and evaluation of chemically induced liver injury. In: Wallace Hayes A (ed) Principles and methods of toxicology, 2nd edn. Raven, New York, pp 399–428Google Scholar
  24. Rahman MA, Dhar DK, Masunaga R, Yamanoi A, Kohno H, Nagasue N (2000) Sulindac and exisulind exhibit a significant antiproliferative effect and induce apoptosis in human hepatocellular carcinoma cell lines. Cancer Res 60:2085–2089Google Scholar
  25. Rumi MA, Sato H, Ishihara S, Kawashima K, Kazumori SHH, Okuyama T, Fukuda R, Nagasue N, Kinoshita Y (2001) Peroxisome proliferator activated receptor gamma ligand induced growth inhibition of human hepatocellular carcinoma. Br J Cancer 84:1640–1647CrossRefGoogle Scholar
  26. Shinya TP, Keisei O, Junji Y, Hiroshi H (1995) Persistent oxidative stress in cancer. FEBS Lett 13:358Google Scholar
  27. Sivaramakrishnan V, Shilpa PN, Kumar VRP, Devarajm SN (2008) Attenuation of N-nitrosodiethylamine-induced hepatocellular carcinogenesis by a novel flavonol Morin. Chem Biol Interact 171:79–88CrossRefGoogle Scholar
  28. Subbaramaiah K, Dannenberg AJ (2003) Cyclooxygenase 2: a molecular target for cancer prevention and treatment. Trends Pharmacol Sci 24:96–102CrossRefGoogle Scholar
  29. Thirunavukkarasu C, Sakthisekaran D (2003) Sodium selenite modulates tumour marker indices in N-nitrosodiethylamine initiated and phenobarbital-promoted rat liver carcinogenesis. Cell Biochem Funct 21:147–153CrossRefGoogle Scholar
  30. Vafa O, Wade M, Kern S (2002) C-Myc can induce DNA damage, increase reactive oxygen species, and mitigate p53 function: a mechanism for oncogene-induced geneticin stability. Mol Cell 9:1031–1044CrossRefGoogle Scholar
  31. Wu T (2006) Cyclooxy genase-2 in hepatocellular carcinoma. Cancer Treat Rev 32:28–44CrossRefGoogle Scholar
  32. Xu-yinga W, Minga L, Xiao-dongb L, Ping H (2009) Hepatoprotective and anti-hepatocarcinogenic effects of glycyrrhizin and matrine. Chem Biol Interact 181:15–19CrossRefGoogle Scholar
  33. Yadav AS, Bhatnagar D (2007) Chemo-preventive effect of staranise in N-nitrosodiethylamine initiated and phenobarbital promoted hepato-carcinogenesis. Chem Biol Interact 169:207–214CrossRefGoogle Scholar
  34. Zhang JF, Liu JJ, Lu MQ, Cai CJ, Yangm Y, Li H, Xu C, Chen GH (2007) Rapamycin inhibits cell growth by induction of apoptosis on hepatocellular carcinoma cells in vitro. Transpl Immunol 17:162–168CrossRefGoogle Scholar

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© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmacology, College of PharmacyJouf UniversitySakakaKingdom of Saudi Arabia
  2. 2.Siddhartha Institute of PharmacyDehradunIndia
  3. 3.Glocal School of PharmacyGlocal UniversitySahranpurIndia
  4. 4.Department of Biochemistry, Faculty of ScienceKing Abdulaziz UniversityJeddahSaudi Arabia

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