Correlation of susceptibility to ortho-aminoazotolueneinduced hepatocarcinogenesis with Car and Ahr signaling pathway activation in mice. N. V. Baginskaya, E. V. Kashina, M. Yu. Shamanina, S. I. Ilnitskaya, V. I. Kaledin, V. A. Mordvinov


Ortho-aminoazotoluene (OAT) is a potent hepatocarcinogen for most strains of mice. It has previously been shown that OAT application activates the aryl hydrocarbon receptor (Ahr) and the constitutive androstane receptor (Car) in the mouse liver. Both of these receptors are directly involved in the process of hepatocarcinogenesis. In this study, we investigated the effect of chronic OAT administration on the mRNA expression levels of Ahr, Car and their target genes Cyp1a1 and Cyp2b10 in the liver of DD/He (DD) and CC57BR/Mv (BR) mouse strains contrasting in sensitivity to hepatocarcinogenesis. The inflammatory response of these strains was also studied. Male mice of both strains received OAT oil solution at the dose of 225 mg/kg body weight four times within two months. Control animals received the equivalent solvent amount. Mice were sacrificed on days 1 and 4 after the last OAT administration. Gene expression levels in the liver were determined by real-time PCR. The inflammatory response was evaluated by serum concentration of tumor necrosis factor alpha (TNF-alpha). In resistant BR mice, OAT induced a pronounced and prolonged increase in Cyp1a1 mRNA, showing primarily Ahr activation, while the DD strain displayed a more pronounced elevation of Cyp2b10 expression, indicative of Car activation. In addition, a strong inflammatory response to OAT was recorded in DD mice but not in BR. It is assumed that the prevalence of Ahr signaling pathway activation over Car signaling pathway activation is a factor of resistance to OAT-induced hepatocarcinogenesis.

About The Authors:

N. V. Baginskaya. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

E. V. Kashina. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

M. Yu. Shamanina. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

S. I. Ilnitskaya. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

V. I. Kaledin. Institute of Cytology and Genetics SB RAS, Russian Federation, Novosibirsk

V. A. Mordvinov. Institute of Cytology and Genetics SB RAS; Institute of Molecular Biology and Biophysics, Russian Federation, Novosibirsk


1. Baginskaya N.V., Il’nitskaya S.I., Nikitenko E.V., Kaledin V.I. Promoting Effect of o-Aminoazotoluene on Hepatocarcinogenesis is Accompanied by the Increase in Inflammatory and Proliferative Processes in Liver Tissue and Decrease in the Concentration of Free Thyroxin in the Blood. Byulleten eksperimentalnoj biologii i meditsini — Bulletin of Experimental Biology and Medicine. 2007;144(12):672-676.

2. Baginskaya N.V., Perepechaeva M.L., Hochenko O.M., Dushkin M.I. The effect of o-aminoazotoluene on PPAR-α, PPAR-γ, RXR-α and CAR genes expression and serum level of IL-1β in mouse strains, contrasting in their susceptibility to chemical hepatocarcinogenesis. Mater. mezhdunar. conf. «Retseptsiya i vnutrikletochnaya signalizatsiya» [Abstr. of the International Conference «Reception and intercellular signaling»]. Pushchino, 2009;1:326-330.

3. Beigneux A.P., Moser A.H., Shigenaga J.K., Grunfeld C., Feingold K.R. Reduction in cytochrome P-450 enzyme expression is associated with repression of CAR (constitutive androstane receptor) and PXR (pregnane X receptor) in mouse liver during the acute phase response. Biochem. Biophys Res. Commun. 2002;293:145-149.

4. Chen C.J., Chen D.S. Interaction of hepatitis B virus, chemical carcinogen, and genetic susceptibility: Multistage hepatocarcinogenesis with multifactorial etiology. Hepatology. 2002;36:1046-1049.

5. Cheung Y.L., Puddicombe S.M., Gray T.J., Ioannides C. Mutagenicity and CYP1A induction by azobenzenes correlates with their carcinogenicity. Carcinogenesis. 1994;15:1257-1263.

6. Fan Y., Boivin G.P., Knudsen E.S., Nebert D.W., Xia Y., Puga A. The aryl hydrocarbon receptor functions as a tumor suppressor of liver carcinogenesis. Cancer. Res. 2010;70:212-220.

7. Feo F., De Miglio M.R., Simile M.M., Muroni M.R., Calvisi D.F., Frau M., Pascale R.M. Hepatocellular carcinoma as a complex polygenic disease. Interpretive analysis of recent developments on genetic predisposition. Biochim. Biophys. Acta. 2006;1765:126-147.

8. Gerbal-Chaloin S., Iankova I., Maurel P., Daujat-Chavanieu M. Nuclear receptors in the cross-talk of drug metabolism and inflammation. Drug Metab. Rev. 2013;45(1):122-144.

9. Glass C.K., Saijo K. Nuclear receptor transrepression pathways that regulate inflammation in macrophages and T cells. Nat. Rev. Immunol. 2010;10:365-376. DOI: 10.1038/nri2748

10. Hanieh H. Toward understanding the role of aryl hydrocarbon receptor in the immune system: current progress and future trends. Biomed. Res. Int. 2014. DOI: 10.1155/2014/520763

11. Huang W., Zhang J., Washington M., Liu J., Parant J.M., Lozano G., Moore D.D. Xenobiotic stress induces hepatomegaly and liver tumors via the nuclear receptor constitutive androstane receptor. Mol. Endocrinol. 2005;19:1646-1653.

12. Kaledin V.I., Il’nitskaia S.I. Metabolism inhibition stimulates, metabolism activation inhibits cancerogenic activity of ortho-aminoazotoluene in mouse liver. Voprosy onkologii — Problems in onkology. 2011;57:216-220.

13. Kaledin V.I., Serova I.A., Semenova L.A. Unequal predisposition to development of spontaneous and induced liver tumors in mice of different strains and their hybrids. Eksperim. onkologiya — Experimental Onkology. 1990;12(4):28-30.

14. Kolaya K.L., Stevenson D.E., Walborg Jr. E.F., Klaunig J.E. Cancer Biology: Dose dependence of phenobarbital promotion of preneoplastic hepatic lesions in F344 rats and B6C3F1 mice: effects on DNA synthesis and apoptosis. Carcinogenesis. 1995;17:947-954.

15. Liedtke C., Trautwein C. The role of TNF and Fas dependent signaling in animal models of inflammatory liver injury and liver cancer. Eur. J. Cell Biol. 2012;91(6/7):582-589. DOI: 10.1016/j.ejcb.2011.10.001

16. Marlowe J.L., Puga A. Aryl hydrocarbon receptor, cell cycle regulation, toxicity, and tumorigenesis. J. Cell Biochem. 2005;96:1174-1184.

17. Mikhailova O.N., Vasyunina E.A., Ovchinnikova L.P., Gulyaeva L.F., Timofeeva O.A., Filipenko M.L., Kaledin V.I. o-Aminoazotoluene does induce the enzymes of its own mutagenic activation in mouse liver. Toxicology. 2005;211:132-138.

18. Miller E.C., Miller J.A. Searches for ultimate chemical carcinogens and their reactions with cellular macromolecules. Cancer. 1981;47(10): 2327-2345.

19. Mitchell K.A., Lockhart C.A., Huang G., Elferink C.J. Sustained aryl hydrocarbon receptor activity attenuates liver regeneration. Mol. Pharmacol. 2006;70(1):163-170.

20. Moennikes O., Loeppen S., Buchmann A., Andersson P., Ittrich C., Poellinger L., Schwarz M.A. A constitutively active dioxin/aryl hydrocarbon receptor promotes hepatocarcinogenesis in mice. Cancer Res. 2004;64:4707-4710.

21. Nakagawa H., Maeda S. Inflammation- and stress-related signaling pathways in hepatocarcinogenesis. World J. Gastroenterol. 2012; 18(31):4071-4081.

22. Nebert D.W., Dalton T.P., Okey A.B., Gonzalez F.J. Role of aryl hydrocarbon receptor-mediated induction of the CYP1 enzymes in environmental toxicity and cancer. J. Biol. Chem. 2004;279: 23847-23850.

23. Patel R.D., Hollingshead B.D., Omiecinski C.J., Perdew G.H. Arylhydrocarbon receptor activation regulates constitutive androstane receptor levels in murine and human liver. Hepatology. 2007;46: 209-218.

24. Petrick J.S., Klaassen C.D. Importance of hepatic induction of constitutive androstane receptor and other transcription factors that regulate xenobiotic metabolism and transport. Drug Metab. Dispos. 2007;35(10):1806-1815.

25. Smetanina M.A., Pakharukova M.Y., Kurinna S.M., Dong B., Hernandez J.P., Moore D.D., Merkulova T.I. Ortho-aminoazotoluene activates mouse constitutive androstane receptor (mCAR) and increases expression of mCAR target genes. Toxicol. Appl. Pharmacol. 2011;255:76-85.

26. Tian Y., Ke S., Denison M.S., Rabson A.B., Gallo M.A. Ah receptor and NF-kappaB interactions, a potential mechanism for dioxin toxicity. J. Biol. Chem. 1999;274:510-515.

27. Timofeeva O.A., Filipenko M.L., Rychkova N.A., Gulyaeva L.F., Lyakhovich V.V. Expression of CYP1A in the mice of strains A/SN and CC57BR, differing in their sensitivity to a hepatocarcinogenic action of o-aminoazotoluene. Biokhimiya — Biochemistry (Moscow), 2000;65(6):718-722.

28. Vogel C.F., Matsumura F. A new cross-talk between the aryl hydrocarbon receptor and RelB, a member of the NF-kappaB family. Biochem. Pharmacol. 2009;77:734-745.

29. Xie W., Yeuh M.F., Radominska-Pandya A., Saini S.P., Negishi Y., Bottroff B.S., Cabrera G.Y., Tukey R.H., Evans R.M. Control of steroid, heme, and carcinogen metabolism by nuclear pregnane X receptor and constitutive androstane receptor. Proc. Natl Acad. Sci. USA. 2003;100:4150-4155.

30. Yamamoto Y., Moore R., Goldsworthy T.L., Negishi M., Maronpot R.R. The orphan nuclear receptor constitutive active androstane receptor is essential for liver tumor promotion by phenobarbital in mice. Cancer Res. 2004;64(20):7197-7200.

31. Zacharova L.Yu., Gulyaeva L.F., Lyakhovich V.V., Mikhailova O.N., Timofeeva O.A., Filipenko M.L., Kaledin V.I. Cytochrome P4501A1 and 1A2 gene expression in the liver of 3-methylcholanthrene- and o-aminoazotoluene-treated mice: a comparison between PAH-responsive and PAH-nonresponsive strains. Toxicol. Sci. 2003;73: 108-113.

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