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Hepatic inflammation-fibrosis-cancer axis in the rat hepatocellular carcinoma induced by diethylnitrosamine

  • Yong-fang Ding
  • Zhen-hui Wu
  • Ying-jie Wei
  • Luan Shu
  • Yun-ru Peng
Original Article – Cancer Research

Abstract

Purpose

Hepatocellular carcinoma (HCC) cases are closely associated with chronic inflammation and fibrosis which is known as hepatic inflammation-fibrosis-cancer (IFC) axis. The aim of this study is to elucidate the development characteristics of the rat HCC model based on IFC axis.

Methods

The diethylnitrosamine (DEN)-induced rat HCC, which presents a stepwise histopathological progression that is similar to human HCC, was used to analyze the features of the different stages (inflammation, fibrosis, cancer). Rats were injected DEN at a dose of 30 mg/kg body weight twice a week for 11 weeks and the animals were observed until week 20. Time series sera and organ samples from the DEN animal model were collected to evaluate the dynamic changes.

Results

It was found that serum biochemical indicators (AST, ALT, ALP, TP, T-BIL, IL-6, TNF-α) from DEN-treated group were higher than that from control group. Fibrosis-related index in serum and live tissue were increased, respectively, from week 4 after DEN treatment. The expression of TGF-β1 and α-SMA in DEN-treated group was higher than that in control group. JAK2/STAT3 signaling was significantly up-regulated in DEN-treated group compared to that in control group. The histological examination confirmed that the hepatocarcinogenesis model was successfully established, and 100% of the animals in the DEN-exposed group developed liver tumors at 20 weeks. According to the pathological changes, the model characterized resulted in three stages: the inflammation stage (week 2–6), the fibrosis stage (week 8–12), and the HCC stage (week 14–20).

Conclusions

The results suggested that the HCC development was associated with IFC axis. The serial progression of hepatocarcinogenesis was according to the sequence of hepatic inflammation, fibrosis and then hepatic tumor.

Keywords

Hepatocellular carcinoma Liver inflammation Liver fibrosis Inflammation-fibrosis-cancer (IFC) axis 

Notes

Compliance with ethical standards

Conflict of interest

The author(s) declare(s) that they have no conflicts of interest to disclose.

Funding

This study was supported by a Grant from the National Natural Science Foundation of China (Nos. 81373888, 81102884).

Ethical approval

This study was approved by the Animal Experimental Ethical Committee of Jiangsu Province Academy of Traditional Chinese Medicine.

References

  1. Aravalli RN, Cressman EN, Steer CJ (2013) Cellular and molecular mechanisms of hepatocellular carcinoma: an update. Arch Toxicol 87:227–247. doi: 10.1007/s00204-012-0931-2 CrossRefPubMedGoogle Scholar
  2. Bissell DM (2001) Chronic liver injury, TGF-β and cancer. Exp Mol Med 33:179–190CrossRefPubMedGoogle Scholar
  3. Borbath I, Stärkel P (2011) Chemoprevention of hepatocellular carcinoma. Proof of concept in animal models. Acta Gastroenterol Belg 74:34–44PubMedGoogle Scholar
  4. Calès P, Boursier J, Chaigneau J et al (2010) Diagnosis of different liver fibrosis characteristics by blood tests in non-alcoholic fatty liver disease. Liver Int 30:1346–1354CrossRefPubMedGoogle Scholar
  5. Costentin CE, Roudot-Thraval F, Zafrani ES et al (2011) Association of caffeine intake and histological features of chronic hepatitis C. J Hepatol 54:1123–1129CrossRefPubMedGoogle Scholar
  6. Cubero FJ (2016) Shutting off inflammation: a novel switch on hepatic stellate cells. Hepatology 63:1086–1089CrossRefPubMedGoogle Scholar
  7. Di Virgilio F (2014) Purinergic receptors in cancer and inflammation: from Rudolph Virchow to Geoff Burnstock. Purinergic signallling, vol 10. Springer, Netherlands, pp 657–658Google Scholar
  8. Ding Y, Peng Y, Li J et al (2013) Gualou Xiebai Decoction prevents myocardial fibrosis by blocking TGF-beta/Smad signalling. J Pharm Pharmacol 65:1373–1381CrossRefPubMedGoogle Scholar
  9. Duan XY, Pan Q, Yan SY et al (2014) High-saturate-fat diet delays initiation of diethylnitrosamine-induced hepatocellular carcinoma. BMC Gastroenterol 14:195. doi: 10.1186/s12876-014-0195-9 CrossRefPubMedPubMedCentralGoogle Scholar
  10. Elsharkawy AM, Mann DA (2007) Nuclear factor-kappaB and the hepatic inflammation-fibrosis-cancer axis. Hepatology 46:590–597.CrossRefPubMedGoogle Scholar
  11. Farazi PA, Depinho RA (2006) Hepatocellular carcinoma pathogenesis: from genes to environment. Nat Rev Cancer 6:674–687CrossRefPubMedGoogle Scholar
  12. Geiger-Maor A, Guedj A, Even-Ram S (2015) Macrophages regulate the systemic response to DNA damage by a cell nonautonomous mechanism. Cancer Res 75:2663–2673. doi: 10.1158/0008-5472.CAN-14-3635 CrossRefPubMedGoogle Scholar
  13. Grivennikov SI, Greten FR, Karin M (2010) Immunity, inflammation, and cancer. Cell 140:883–899CrossRefPubMedPubMedCentralGoogle Scholar
  14. Hernandez–Gea V, Toffanin S, Friedman SL et al (2013) Role of the microenvironment in the pathogenesis and treatment of hepatocellular carcinoma. Gastroenterology 144:512–527CrossRefGoogle Scholar
  15. Kishida N, Matsuda S, Itano O (2016) Development of a novel mouse model of hepatocellular carcinoma with nonalcoholic steatohepatitis using a high-fat, choline-deficient diet and intraperitoneal injection of diethylnitrosamine. BMC Gastroenterol 16:61. doi: 10.1186/s12876-016-0477-5 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Knodell RG, Ishak KG, Black WC et al (1981) Forumlation and application of a numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis. Hepatology 1:431–435CrossRefPubMedGoogle Scholar
  17. Kong L, Zhou Y, Bu H et al (2016) Deletion of interleukin-6 in monocytes/macrophages suppresses the initiation of hepatocellular carcinoma in mice. J Exp Clin Cancer Res 35:131. doi: 10.1186/s13046-016-0412-1 CrossRefPubMedPubMedCentralGoogle Scholar
  18. Li J, Sun Z, Zhang J et al (2011) A dual-targeting anticancer approach: soil and seed principle. Radiology 260:799–807CrossRefPubMedGoogle Scholar
  19. Li C, Bi X, Huang Y et al (2014) Variants identified by hepatocellular carcinoma and chronic hepatitis B virus infection susceptibility GWAS associated with survival in HBV-related hepatocellular carcinoma. PloS One 9(7):e101586. doi: 10.1371/journal.pone.0101586 CrossRefPubMedPubMedCentralGoogle Scholar
  20. Ma W, Sze KM, Chan LK et al (2016) RhoE/ROCK regulates chemoresistance through NF-ΚB/IL-6/STAT3 signaling in hepatocellular carcinoma. Oncotarget 27:41445–41459. doi: 10.18632/oncotarget.9441 Google Scholar
  21. Peng Y, Ding Y (2015) Pharmacokinetics and tissue distribution study of caudatin in normal and diethylnitrosamine-induced hepatocellular carcinoma model rats. Molecules 20:4225–4237CrossRefPubMedGoogle Scholar
  22. Santos NP, Colaco A, da Costa R M G (2014) N-diethylnitrosamine mouse hepatotoxicity: time-related effects on histology and oxidative stress. Exp Toxicol Pathol 66:429–436CrossRefGoogle Scholar
  23. Tachi Y, Hirai T, Miyata A et al (2015) Progressive fibrosis significantly correlates with hepatocellular carcinoma in patients with a sustained virological response. Hepatol Res 45:238–246. doi: 10.1111/hepr.12331 CrossRefPubMedGoogle Scholar
  24. Takahashi Y, Fukusato T (2014) Histopathology of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. World J Gastroenterol 20:15539–15548. doi: 10.3748/wjg.v20.i42.15539 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Tyler PD, Mcdevitt JL, Sheu AY et al (2013) Seven-tesla magnetic resonance imaging accurately quantifies intratumoral uptake of therapeutic nanoparticles in the McA rat model of hepatocellular carcinoma: preclinical study in a rodent model. Investig Radiol 49:87–92CrossRefGoogle Scholar
  26. Uehara T, Pogribny I P, Rusyn I (2014) The DEN and CCl4-induced mouse model of fibrosis and inflammation-associated hepatocellular carcinoma. Curr Protoc Pharmacol 66:14.30.1–14.30.10Google Scholar
  27. Wong WS, Janssen H L A (2015) Can we use HCC risk scores to individualize surveillance in chronic hepatitis B infection? J Hepatol 63:722–732CrossRefPubMedGoogle Scholar
  28. Zeng J, Huang X, Zhou L et al (2015) Metaboomics identifies biomarker pattern for early diagnosis of hepatocellular carcinoma: from diethylnitrosamine treated rats to patients. Sci Rep 5:16101. doi: 10.1038/srep16101.CrossRefPubMedPubMedCentralGoogle Scholar
  29. Zhao H, Guo Y, Han R et al (2015) A novel anti-cancer agent Icaritin suppresses hepatocellular carcinoma initiation and malignant growth through the IL-6/Jak2/Stat3 pathway. Oncotarget 31:31927–31943. doi: 10.18632/oncotarget.5578 Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Yong-fang Ding
    • 1
    • 2
  • Zhen-hui Wu
    • 3
  • Ying-jie Wei
    • 1
    • 2
  • Luan Shu
    • 1
    • 2
  • Yun-ru Peng
    • 1
    • 2
  1. 1.Affiliated Hospital of Integrated Traditional Chinese and Western MedicineNanjing University of Chinese MedicineNanjingPeople’s Republic of China
  2. 2.Jiangsu Province Academy of Traditional Chinese MedicineNanjingPeople’s Republic of China
  3. 3.Nanjing University of Chinese MedicineNanjingPeople’s Republic of China

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