Modulatory effects of rutin on the expression of cytochrome P450s and antioxidant enzymes in human hepatoma cells

Open access


Expression of a drug and xenobiotic metabolizing enzymes, cytochrome P450s (CYPs), and antioxidant enzymes can be modulated by various factors. The flavonoid rutin was investigated for its anti-carcinogen and protective effects as well as modulatory action on CYPs and phase II enzymes in human hepatocellular carcinoma cells. Rutin inhibited proliferation of HEPG2 cells in a dose-dependent manner with the IC50 value of 52.7 μmol L-1 and invasion of HEPG2 cells (21.6 %, p = 0.0018) and colony formation of those invaded cells (57.4 %, p < 0.0001). Rutin treatment also significantly increased early/late-stage apoptosis in HEPG2 cells (28.9 %, p < 0.001). Treatment by rutin significantly inhibited protein expressions of cytochrome P450-dependent CYP3A4 (75.3 %, p < 0.0001), elevated CYP1A1 enzymes (1.7-fold, p = 0.0084) and increased protein expressions of antioxidant and phase II reaction catalyzing enzymes, NQO1 (2.42-fold, p < 0.0001) and GSTP1 (2.03-fold, p < 0.0001). Besides, rutin treatment significantly inhibited mRNA expression of CYP3A4 (73.2 %, p=0.0014). Also, CYP1A1, NQO1 and GSTP1 mRNA expressions were significantly increased 2.77-fold (p = 0.029), 4.85- fold (p = 0.0051) and 9.84-fold (p < 0.0001), respectively.

1. L. A. Torre, F. Bray, R. L. Siegel, J. Ferlay, J. Lortet-Tieulent and A. Jemal, Global cancer statistics, 2012, Cancer J. Clin. 65 (2015) 87-108; DOI: 10.3322/caac.21262.

2. M. Balbi, V. Donadon, M. Ghersetti, S. Grazioli, G. D. Valentina, R. Gardenal, M. D. Mas, P. Casarin, G. Zanette, C. Miranda and P. Cimarosti, Alcohol and HCV chronic infection are risk cofactors of type 2 diabetes mellitus for hepatocellular carcinoma in Italy, Int. J. Environ. Res. Public Health 7 (2010) 1366-1378; DOI: 10.3390/ijerph7041366.

3. X. Zhang and H. G. Ding, Key role of hepatitis B virus mutation in chronic hepatitis B development to hepatocellular carcinoma, World J. Hepatol. 7 (2015) 1282-1286; DOI: 10.4254/wjh.v7.i9.1282.

4. P. C. Hollman and M. B. Katan, Absorption, metabolism and health effects of dietary flavonoids in man, Biomed. Pharmacother. 51 (1997) 305-310; DOI: 10.1016/S0753-3322(97)88045-6.

5. E. Middleton, C. Kandaswami and T. C. Theoharides, The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer, Pharmacol. Rev. 52 (2000) 673-751.

6. G. Williamson and C. Manach, Bioavailability and bioefficacy of polyphenols in humans. II. Review of 93 intervention studies, Am. J. Clin. Nutr. 81 (2005) 243S-255S.

7. B. A. Graf, C. Ameho, G. G. Dolnikowski, P. E. Milbury, C. Y. Chen and J. B. Blumberg, Rat gastrointestinal tissues metabolize quercetin, J. Nutr. 136 (2006) 39-44.

8. S. Scholz and G. Williamson, Interactions affecting the bioavailability of dietary polyphenols in vivo, Int. J. Vitam. Nutr.Res. 77 (2007) 224-235; DOI: 10.1024/0300-9831.77.3.224.

9. V. A. Kostyuk, A. I. Potapovich, T. V. Kostyuk and M. G. Cherian, Metal complexes of dietary flavonoids: evaluation of radical scavenger properties and protective activity against oxidative stress in vivo, Cell. Mol. Biol. (Noisy-le-grand) 53 (2007) 62-69; DOI: 10.1170/T776.

10. K. Song, J. Y. Na, S. Kim and J. Kwon, Rutin upregulates neurotrophic factors resulting in attenuation of ethanol-induced oxidative stress in HT22 hippocampal neuronal cells, J. Sci. Food Agric. 95 (2015) 2117-2123; DOI: 10.1002/jsfa.6927.

11. B. L. Santos, A. R. Silva, B. P. Pitanga, C. S. Sousa, M. S. Grangeiro, B. O. Fragomeni, P. L. Coelho, M. N. Oliveira, N. J. Menezes-Filho, M. F. Costa, R. S. El-Bachá, E. S. Velozo, G. P. Sampaio, S. M. Freire, M. Tardy and S. L. Costa, Antiproliferative, proapoptotic and morphogenic effects of the flavonoid rutin on human glioblastoma cells, Food Chem. 127 (2011) 404-411; DOI: 10.1016/j.foodchem. 2010.12.13.

12. N. Ihme, H. Kiesewetter, F. Jung, K. H. Hoffmann, A. Birk, A. Müller and K. I. Grützner, Leg oedema protection from a buckwheat herb tea in patients with chronic venous insufficiency: a single- centre, randomised, double-blind, placebo-controlled clinical trial, Eur. J. Clin. Pharmacol. 50 (1996) 443-447; DOI: 10.1007/s002280050138.

13. S. Okonogi, R. Khonkarn, S. Mankhetkorn, F. M. Unger and H. Viernstein, Antioxidant activity and cytotoxicity of Cyrtosperma johnstonii extracts on drug sensitive and resistant leukemia and small cell lung carcinoma cells, Pharm. Biol. 51 (2013) 329-338; DOI: 10.3109/13880209.2012.729064.

14. H. J. You, H. J. Ahn and G. E. Ji, Transformation of rutin to antiproliferative quercetin-3-glucoside by Aspergillus niger, J. Agric. Food Chem. 58 (2010) 10886-10892; DOI: 10.1021/jf102871g.

15. F. P. Guengerich, Cytochrome P-450 3A4: regulation and role in drug metabolism, Annu. Rev. Pharmacol. Toxicol. 39 (1999) 1-17; DOI: 10.1146/annurev.pharmtox.39.1.1.

16. U. M. Zanger and M. Schwab, Cytochrome P450 enzymes in drug metabolism: regulation of gene expression, enzyme activities, and impact of genetic variation, Pharmacol. Ther. 138 (2013) 103-141; DOI: 10.1016/j.pharmthera.2012.12.007.

17. K. Ohkura, Y. Kawaguchi, Y. Watanabe, Y. Masubuchi, Y. Shinohara and H. Hori, Flexible structure of cytochrome P450: promiscuity of ligand binding in the CYP3A4 heme pocket, Anticancer Res. 29 (2009) 935-942.

18. E. Arinç, D. Yilmaz and A. Bozcaarmutlu, Mechanism of inhibition of CYP1A1 and glutathione S-transferase activities in fish liver by quercetin, resveratrol, naringenin, hesperidin, and rutin, Nutr. Cancer 67 (2015) 137-144; DOI: 10.1080/01635581.2015.965335.

19. T. Fujimura, S. Takahashi, T. Urano, T. Tanaka, W. Zhang, K. Azuma, K. Takayama, D. Obinata, T. Murata, K. Horie-Inoue, T. Kodama, Y. Ouchi, Y. Homma and S. Inoue, Clinical significance of steroid and xenobiotic receptor and its targeted gene CYP3A4 in human prostate cancer, Cancer Sci. 103 (2012) 176-180; DOI: 10.1111/j.1349-7006.2011.02143.x.

20. J. C. Stevens, R. N. Hines, C. Gu, S. B. Koukouritaki, J. R. Manro, P. J. Tandler and M. J. Zaya, Developmental expression of the major human hepatic CYP3A enzymes, J. Pharmacol. Exp. Ther. 307 (2003) 573-582; DOI: 10.1124/jpet.103.054841.

21. H. P. Ciolino, P. J. Daschner and G. C. Yeh, Dietary flavonols quercetin and kaempferol are ligands of the aryl hydrocarbon receptor that affect CYP1A1 transcription differentially, Biochem. J. 340 (1999) 715-722; DOI: 10.1042/bj3400715.

22. R. Vrba, R. Aujeský and C. Neoral, Results of limited resection of carcinoma of the gastroesophageal junction, Rozhl. Chir. 91 (2012) 368-372.

23. Mohammadi-Bardbori, J. Bengtsson, U. Rannug, A. Rannug and E. Wincent, Quercetin, resveratrol, and curcumin are indirect activators of the aryl hydrocarbon receptor (AHR), Chem. Res. Toxicol. 25 (2012) 1878-1884; DOI: 10.1021/tx300169e.

24. L. Rongrui, H. Na, L. Zongfang, J. Fanpu and J. Shiwen, Epigenetic mechanism involved in the HBV/HCV-related hepatocellular carcinoma tumorigenesis, Curr. Pharm. Des. 20 (2014) 1715-1725; DOI: 10.2174/13816128113199990533.

25. M. Kiran, Y. K. Chawla and J. Kaur, Methylation profiling of tumor suppressor genes and oncogenes in hepatitis virus-related hepatocellular carcinoma in northern India, Cancer Genet. Cytogenet. 195 (2009) 112-119; DOI: 10.1016/j.cancergencyto.2009.06.021.

26. D. Ross, J. K. Kepa, S. L. Winski, H. D. Beall, A. Anwar and D. Siegel, NAD(P)H:quinone oxidoreductase 1 (NQO1): chemoprotection, bioactivation, gene regulation and genetic polymorphisms, Chem. Biol. Interact. 129 (2000) 77-97; DOI: 10.1016/S0009-2797(00)00199-X.

27. P. Joseph, T. Xie, Y. Xu and A. K. Jaiswal, NAD(P)H:quinone oxidoreductase1 (DT-diaphorase): expression, regulation, and role in cancer, Oncol. Res. 6 (1994) 525-532.

28. E. A. Rosvold, K. A. McGlynn, E. D. Lustbader and K. H. Buetow, Identification of an NAD(P) H:quinone oxidoreductase polymorphism and its association with lung cancer and smoking, Pharmacogenetics 5 (1995) 199-206.

29. Y. Fan, D. Hu, B. Feng and W. Wang, The NQO1 C609T polymorphism and hepatocellular carcinoma risk, Tumour Biol. 35 (2014) 7343-7350; DOI: 10.1007/s13277-014-1712-8.

30. H. Akkiz, S. Bayram, A. Bekar, E. Akgöllü, Y. Ülger, B. Y. Kaya, M. Sandikçi and B. Özdil, No association of NAD(P)H: quinone oxidoreductase 1 (NQO1) C609T polymorphism and risk of hepatocellular carcinoma development in Turkish subjects, Asian Pac. J. Cancer Prev. 11 (2010) 1051-1058.

31. M. Belinsky and A. K. Jaiswal, NAD(P)H:quinone oxidoreductase1 (DT-diaphorase) expression in normal and tumor tissues, Cancer Metastasis Rev. 12 (1993) 103-117; DOI: 10.1007/BF00689804

Acta Pharmaceutica

The Journal of Croatian Pharmaceutical Society

Journal Information

IMPACT FACTOR 2017: 1.071
5-year IMPACT FACTOR: 1.623

CiteScore 2017: 1.46

SCImago Journal Rank (SJR) 2017: 0.362
Source Normalized Impact per Paper (SNIP) 2017: 0.642

Cited By


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 192 192 33
PDF Downloads 105 105 20