Abstract
Hepatocellular carcinoma (HCC), the most frequent form of liver cancer, is among the top fatal malignancies worldwide. The therapeutic options for patients with advanced HCC are limited and poorly efficient. Therefore, it is critical that we identify and develop novel curative strategies in order to address this major health issue. For this, it is essential to gain mechanistic understanding of HCC pathogenesis as clues to therapeutic intervention. Over the last decades, a wide range of HCC models have been developed, aiming to achieve the goal. In particular, animal models and in vitro systems that mimic the major characteristics of the human HCC have emerged to enable functional assessment of candidate therapeutic targets in liver cancer. However, each model recapitulates limited aspects of the whole pathogenesis of the human disease. Nevertheless, combined analysis of multiple different models can collectively provide clinically relevant insights into the complex disease mechanisms. Here we summarize the major approaches of experimental modeling in HCC and their strengths and limitations.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Dhanasekaran R, Limaye A, Cabrera R. Hepatocellular carcinoma: current trends in worldwide epidemiology, risk factors, diagnosis, and therapeutics. Hepat Med. 2012;4:19–37. https://doi.org/10.2147/HMER.S16316.
Llovet JM, Zucman-Rossi J, Pikarsky E, Sangro B, Schwartz M, Sherman M, et al. Hepatocellular carcinoma. Nat Rev Dis Primers. 2016;2:16018. https://doi.org/10.1038/nrdp.2016.18.
Tang ZY, Ye SL, Liu YK, Qin LX, Sun HC, Ye QH, et al. A decade’s studies on metastasis of hepatocellular carcinoma. J Cancer Res Clin Oncol. 2004;130(4):187–96. https://doi.org/10.1007/s00432-003-0511-1.
Portolani N, Coniglio A, Ghidoni S, Giovanelli M, Benetti A, Tiberio GA, et al. Early and late recurrence after liver resection for hepatocellular carcinoma: prognostic and therapeutic implications. Ann Surg. 2006;243(2):229–35. https://doi.org/10.1097/01.sla.0000197706.21803.a1.
Dimitroulis D, Damaskos C, Valsami S, Davakis S, Garmpis N, Spartalis E, et al. From diagnosis to treatment of hepatocellular carcinoma: an epidemic problem for both developed and developing world. World J Gastroenterol. 2017;23(29):5282–94. https://doi.org/10.3748/wjg.v23.i29.5282.
Mittal S, El-Serag HB. Epidemiology of hepatocellular carcinoma: consider the population. J Clin Gastroenterol. 2013;47(Suppl):S2–6. https://doi.org/10.1097/MCG.0b013e3182872f29.
Gillet JP, Varma S, Gottesman MM. The clinical relevance of cancer cell lines. J Natl Cancer Inst. 2013;105(7):452–8. https://doi.org/10.1093/jnci/djt007.
Collection AATC. Hep G2 [HEPG2] (ATCC® HB-8065™). ATCC. 2016. https://www.atcc.org/products/all/HB-8065.aspx#characteristics. 2018.
Lopez-Terrada D, Cheung SW, Finegold MJ, Knowles BB. Hep G2 is a hepatoblastoma-derived cell line. Hum Pathol. 2009;40(10):1512–5. https://doi.org/10.1016/j.humpath.2009.07.003.
Bank JC. JCRB0403 – Huh-7. National Institutes of Biomedical Innovation, Health and Nutrition – Japan. 2015. http://cellbank.nibiohn.go.jp/~cellbank/en/search_res_det.cgi?ID=385.2018.
Sainz B Jr, TenCate V, Uprichard SL. Three-dimensional Huh7 cell culture system for the study of hepatitis C virus infection. Virol J. 2009;6:103. https://doi.org/10.1186/1743-422X-6-103.
Fang C, Yi Z, Liu F, Lan S, Wang J, Lu H, et al. Proteome analysis of human liver carcinoma Huh7 cells harboring hepatitis C virus subgenomic replicon. Proteomics. 2006;6(2):519–27. https://doi.org/10.1002/pmic.200500233.
Ali N, Allam H, May R, Sureban SM, Bronze MS, Bader T, et al. Hepatitis C virus-induced cancer stem cell-like signatures in cell culture and murine tumor xenografts. J Virol. 2011;85(23):12292–303. https://doi.org/10.1128/JVI.05920-11.
Bressac B, Galvin KM, Liang TJ, Isselbacher KJ, Wands JR, Ozturk M. Abnormal structure and expression of p53 gene in human hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1990;87(5):1973–7.
Rebouissou S, Zucman-Rossi J, Moreau R, Qiu Z, Hui L. Note of caution: contaminations of hepatocellular cell lines. J Hepatol. 2017;67(5):896–7. https://doi.org/10.1016/j.jhep.2017.08.002.
Ramboer E, Vanhaecke T, Rogiers V, Vinken M. Immortalized human hepatic cell lines for in vitro testing and research purposes. Methods Mol Biol. 2015;1250:53–76. https://doi.org/10.1007/978-1-4939-2074-7_4.
Arellanes-Robledo J, Hernández C, Camacho J, Pérez-Carreón JI, editors. In vitro models of HCC, Chapter 42. In: Liver pathophysiology: Academic Press; 2017. https://www.elsevier.com/books/liver-pathophysiology/muriel/978-0-12-804274-8.
Fatehullah A, Tan SH, Barker N. Organoids as an in vitro model of human development and disease. Nat Cell Biol. 2016;18(3):246–54. https://doi.org/10.1038/ncb3312.
Clevers H. Modeling development and disease with organoids. Cell. 2016;165(7):1586–97. https://doi.org/10.1016/j.cell.2016.05.082.
Takebe T, Sekine K, Enomura M, Koike H, Kimura M, Ogaeri T, et al. Vascularized and functional human liver from an iPSC-derived organ bud transplant. Nature. 2013;499(7459):481–4. https://doi.org/10.1038/nature12271.
Huch M, Gehart H, van Boxtel R, Hamer K, Blokzijl F, Verstegen MM, et al. Long-term culture of genome-stable bipotent stem cells from adult human liver. Cell. 2015;160(1–2):299–312. https://doi.org/10.1016/j.cell.2014.11.050.
Broutier L, Andersson-Rolf A, Hindley CJ, Boj SF, Clevers H, Koo BK, et al. Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation. Nat Protoc. 2016;11(9):1724–43. https://doi.org/10.1038/nprot.2016.097.
Huch M, Dorrell C, Boj SF, van Es JH, Li VS, van de Wetering M, et al. In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature. 2013;494(7436):247–50. https://doi.org/10.1038/nature11826.
Broutier L, Mastrogiovanni G, Verstegen MM, Francies HE, Gavarro LM, Bradshaw CR, et al. Human primary liver cancer-derived organoid cultures for disease modeling and drug screening. Nat Med. 2017;23(12):1424–35. https://doi.org/10.1038/nm.4438.
He L, Tian DA, Li PY, He XX. Mouse models of liver cancer: progress and recommendations. Oncotarget. 2015;6(27):23306–22. https://doi.org/10.18632/oncotarget.4202.
Price JE. Xenograft models in immunodeficient animals: I. Nude mice: spontaneous and experimental metastasis models. Methods Mol Med. 2001;58:205–13. https://doi.org/10.1385/1-59259-137-X:205.
Morton CL, Houghton PJ. Establishment of human tumor xenografts in immunodeficient mice. Nat Protoc. 2007;2(2):247–50. https://doi.org/10.1038/nprot.2007.25.
Richmond A, Su Y. Mouse xenograft models vs GEM models for human cancer therapeutics. Dis Model Mech. 2008;1(2–3):78–82. https://doi.org/10.1242/dmm.000976.
Heindryckx F, Colle I, Van Vlierberghe H. Experimental mouse models for hepatocellular carcinoma research. Int J Exp Pathol. 2009;90(4):367–86. https://doi.org/10.1111/j.1365-2613.2009.00656.x.
Santos NP, Colaco AA, Oliveira PA. Animal models as a tool in hepatocellular carcinoma research: a review. Tumour Biol. 2017;39(3):1010428317695923. https://doi.org/10.1177/1010428317695923.
Killion JJ, Radinsky R, Fidler IJ. Orthotopic models are necessary to predict therapy of transplantable tumors in mice. Cancer Metastasis Rev. 1998;17(3):279–84.
El-Khoueiry AB, Sangro B, Yau T, Crocenzi TS, Kudo M, Hsu C, et al. Nivolumab in patients with advanced hepatocellular carcinoma (CheckMate 040): an open-label, non-comparative, phase 1/2 dose escalation and expansion trial. Lancet. 2017;389(10088):2492–502. https://doi.org/10.1016/S0140-6736(17)31046-2.
Zhu AX, Finn RS, Edeline J, Cattan S, Ogasawara S, Palmer D, et al. Pembrolizumab in patients with advanced hepatocellular carcinoma previously treated with sorafenib (KEYNOTE-224): a non-randomised, open-label phase 2 trial. Lancet Oncol. 2018; https://doi.org/10.1016/S1470-2045(18)30351-6.
Zimmerman HJ, Lewis JH. Chemical- and toxin-induced hepatotoxicity. Gastroenterol Clin N Am. 1995;24(4):1027–45.
Lee WM. Drug-induced hepatotoxicity. N Engl J Med. 2003;349(5):474–85. https://doi.org/10.1056/NEJMra021844.
Zhang YJ. Interactions of chemical carcinogens and genetic variation in hepatocellular carcinoma. World J Hepatol. 2010;2(3):94–102. https://doi.org/10.4254/wjh.v2.i3.94.
Heath JC. The production of malignant tumours by cobalt in the rat. Br J Cancer. 1956;10(4):668–73.
Verna L, Whysner J, Williams GM. N-nitrosodiethylamine mechanistic data and risk assessment: bioactivation, DNA-adduct formation, mutagenicity, and tumor initiation. Pharmacol Ther. 1996;71(1–2):57–81.
Qi Y, Chen X, Chan CY, Li D, Yuan C, Yu F, et al. Two-dimensional differential gel electrophoresis/analysis of diethylnitrosamine induced rat hepatocellular carcinoma. Int J Cancer. 2008;122(12):2682–8. https://doi.org/10.1002/ijc.23464.
Kolaja KL, Klaunig JE. Vitamin E modulation of hepatic focal lesion growth in mice. Toxicol Appl Pharmacol. 1997;143(2):380–7.
Tolba R, Kraus T, Liedtke C, Schwarz M, Weiskirchen R. Diethylnitrosamine (DEN)-induced carcinogenic liver injury in mice. Lab Anim. 2015;49(1 Suppl):59–69. https://doi.org/10.1177/0023677215570086.
Maronpot RR. Biological basis of differential susceptibility to hepatocarcinogenesis among mouse strains. J Toxicol Pathol. 2009;22(1):11–33. https://doi.org/10.1293/tox.22.11.
Nakatani T, Roy G, Fujimoto N, Asahara T, Ito A. Sex hormone dependency of diethylnitrosamine-induced liver tumors in mice and chemoprevention by leuprorelin. Jpn J Cancer Res. 2001;92(3):249–56.
Naugler WE, Sakurai T, Kim S, Maeda S, Kim K, Elsharkawy AM, et al. Gender disparity in liver cancer due to sex differences in MyD88-dependent IL-6 production. Science. 2007;317(5834):121–4. https://doi.org/10.1126/science.1140485.
Connor F, Rayner TF, Aitken SJ, Feig C, Lukk M, Santoyo-Lopez J, et al. Mutational landscape of a chemically-induced mouse model of liver cancer. J Hepatol. 2018; https://doi.org/10.1016/j.jhep.2018.06.009.
Waxman DJ, Azaroff L. Phenobarbital induction of cytochrome P-450 gene expression. Biochem J. 1992;281(Pt 3):577–92.
Watson RE, Goodman JI. Effects of phenobarbital on DNA methylation in GC-rich regions of hepatic DNA from mice that exhibit different levels of susceptibility to liver tumorigenesis. Toxicol Sci. 2002;68(1):51–8.
Gourama H, Bullerman LB. Aspergillus flavus and Aspergillus parasiticus: Aflatoxigenic fungi of concern in foods and feeds: a review. J Food Prot. 1995;58(12):1395–404.
Liu Y, Wu F. Global burden of aflatoxin-induced hepatocellular carcinoma: a risk assessment. Environ Health Perspect. 2010;118(6):818–24. https://doi.org/10.1289/ehp.0901388.
Guengerich FP, Johnson WW, Shimada T, Ueng YF, Yamazaki H, Langouet S. Activation and detoxication of aflatoxin B1. Mutat Res. 1998;402(1–2):121–8.
Hamid AS, Tesfamariam IG, Zhang Y, Zhang ZG. Aflatoxin B1-induced hepatocellular carcinoma in developing countries: geographical distribution, mechanism of action and prevention. Oncol Lett. 2013;5(4):1087–92. https://doi.org/10.3892/ol.2013.1169.
Mace K, Aguilar F, Wang JS, Vautravers P, Gomez-Lechon M, Gonzalez FJ, et al. Aflatoxin B1-induced DNA adduct formation and p53 mutations in CYP450-expressing human liver cell lines. Carcinogenesis. 1997;18(7):1291–7.
McGlynn KA, Hunter K, LeVoyer T, Roush J, Wise P, Michielli RA, et al. Susceptibility to aflatoxin B1-related primary hepatocellular carcinoma in mice and humans. Cancer Res. 2003;63(15):4594–601.
Chu YJ, Yang HI, Wu HC, Lee MH, Liu J, Wang LY, et al. Aflatoxin B1 exposure increases the risk of hepatocellular carcinoma associated with hepatitis C virus infection or alcohol consumption. Eur J Cancer. 2018;94:37–46. https://doi.org/10.1016/j.ejca.2018.02.010.
Kew MC. Synergistic interaction between aflatoxin B1 and hepatitis B virus in hepatocarcinogenesis. Liver Int. 2003;23(6):405–9.
Boll M, Weber LW, Becker E, Stampfl A. Mechanism of carbon tetrachloride-induced hepatotoxicity. Hepatocellular damage by reactive carbon tetrachloride metabolites. Z Naturforsch C. 2001;56(7–8):649–59.
Bhathal PS, Rose NR, Mackay IR, Whittingham S. Strain differences in mice in carbon tetrachloride-induced liver injury. Br J Exp Pathol. 1983;64(5):524–33.
Dapito DH, Mencin A, Gwak GY, Pradere JP, Jang MK, Mederacke I, et al. Promotion of hepatocellular carcinoma by the intestinal microbiota and TLR4. Cancer Cell. 2012;21(4):504–16. https://doi.org/10.1016/j.ccr.2012.02.007.
Fitzhugh OG, Nelson AA. Liver tumors in rats fed thiourea or thioacetamide. Science. 1948;108(2814):626–8. https://doi.org/10.1126/science.108.2814.626.
Koen YM, Sarma D, Hajovsky H, Galeva NA, Williams TD, Staudinger JL, et al. Protein targets of thioacetamide metabolites in rat hepatocytes. Chem Res Toxicol. 2013;26(4):564–74. https://doi.org/10.1021/tx400001x.
Martinez AK, Maroni L, Marzioni M, Ahmed ST, Milad M, Ray D, et al. Mouse models of liver fibrosis mimic human liver fibrosis of different etiologies. Curr Pathobiol Rep. 2014;2(4):143–53. https://doi.org/10.1007/s40139-014-0050-2.
Li X, Benjamin IS, Alexander B. Reproducible production of thioacetamide-induced macronodular cirrhosis in the rat with no mortality. J Hepatol. 2002;36(4):488–93.
Zeisel SH, da Costa KA. Choline: an essential nutrient for public health. Nutr Rev. 2009;67(11):615–23. https://doi.org/10.1111/j.1753-4887.2009.00246.x.
Corbin KD, Zeisel SH. Choline metabolism provides novel insights into nonalcoholic fatty liver disease and its progression. Curr Opin Gastroenterol. 2012;28(2):159–65. https://doi.org/10.1097/MOG.0b013e32834e7b4b.
Chandar N, Lombardi B. Liver cell proliferation and incidence of hepatocellular carcinomas in rats fed consecutively a choline-devoid and a choline-supplemented diet. Carcinogenesis. 1988;9(2):259–63.
Ikawa-Yoshida A, Matsuo S, Kato A, Ohmori Y, Higashida A, Kaneko E, et al. Hepatocellular carcinoma in a mouse model fed a choline-deficient, L-amino acid-defined, high-fat diet. Int J Exp Pathol. 2017;98(4):221–33. https://doi.org/10.1111/iep.12240.
Kishida N, Matsuda S, Itano O, Shinoda M, Kitago M, Yagi H, et al. 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. 2016;16(1):61. https://doi.org/10.1186/s12876-016-0477-5.
Tsuchida T, Lee YA, Fujiwara N, Ybanez M, Allen B, Martins S, et al. A simple diet- and chemical-induced murine NASH model with rapid progression of steatohepatitis, fibrosis and liver cancer. J Hepatol. 2018;69(2):385–95. https://doi.org/10.1016/j.jhep.2018.03.011.
Hill-Baskin AE, Markiewski MM, Buchner DA, Shao H, DeSantis D, Hsiao G, et al. Diet-induced hepatocellular carcinoma in genetically predisposed mice. Hum Mol Genet. 2009;18(16):2975–88. https://doi.org/10.1093/hmg/ddp236.
Borbath I, Horsmans Y. The role of PPARgamma in hepatocellular carcinoma. PPAR Res. 2008;2008:209520. https://doi.org/10.1155/2008/209520.
Hsu HT, Chi CW. Emerging role of the peroxisome proliferator-activated receptor-gamma in hepatocellular carcinoma. J Hepatocell Carcinoma. 2014;1:127–35. https://doi.org/10.2147/JHC.S48512.
Kersten K, de Visser KE, van Miltenburg MH, Jonkers J. Genetically engineered mouse models in oncology research and cancer medicine. EMBO Mol Med. 2017;9(2):137–53. https://doi.org/10.15252/emmm.201606857.
Lewandoski M. Conditional control of gene expression in the mouse. Nat Rev Genet. 2001;2(10):743–55. https://doi.org/10.1038/35093537.
Bouabe H, Okkenhaug K. Gene targeting in mice: a review. Methods Mol Biol. 2013;1064:315–36. https://doi.org/10.1007/978-1-62703-601-6_23.
El-Serag HB. Epidemiology of viral hepatitis and hepatocellular carcinoma. Gastroenterology. 2012;142(6):1264–73 e1. https://doi.org/10.1053/j.gastro.2011.12.061.
Ott JJ, Stevens GA, Groeger J, Wiersma ST. Global epidemiology of hepatitis B virus infection: new estimates of age-specific HBsAg seroprevalence and endemicity. Vaccine. 2012;30(12):2212–9. https://doi.org/10.1016/j.vaccine.2011.12.116.
Levrero M, Zucman-Rossi J. Mechanisms of HBV-induced hepatocellular carcinoma. J Hepatol. 2016;64(1 Suppl):S84–S101. https://doi.org/10.1016/j.jhep.2016.02.021.
Wang Y, Cui F, Lv Y, Li C, Xu X, Deng C, et al. HBsAg and HBx knocked into the p21 locus causes hepatocellular carcinoma in mice. Hepatology. 2004;39(2):318–24. https://doi.org/10.1002/hep.20076.
Wang HC, Huang W, Lai MD, Su IJ. Hepatitis B virus pre-S mutants, endoplasmic reticulum stress and hepatocarcinogenesis. Cancer Sci. 2006;97(8):683–8. https://doi.org/10.1111/j.1349-7006.2006.00235.x.
Seifer M, Hohne M, Schaefer S, Gerlich WH. In vitro tumorigenicity of hepatitis B virus DNA and HBx protein. J Hepatol. 1991;13(Suppl 4):S61–5.
Koike K. Hepatitis B virus HBx gene and hepatocarcinogenesis. Intervirology. 1995;38(3–4):134–42. https://doi.org/10.1159/000150424.
Zampino R, Marrone A, Restivo L, Guerrera B, Sellitto A, Rinaldi L, et al. Chronic HCV infection and inflammation: clinical impact on hepatic and extra-hepatic manifestations. World J Hepatol. 2013;5(10):528–40. https://doi.org/10.4254/wjh.v5.i10.528.
Lerat H, Honda M, Beard MR, Loesch K, Sun J, Yang Y, et al. Steatosis and liver cancer in transgenic mice expressing the structural and nonstructural proteins of hepatitis C virus. Gastroenterology. 2002;122(2):352–65.
Koike K. Molecular basis of hepatitis C virus-associated hepatocarcinogenesis: lessons from animal model studies. Clin Gastroenterol Hepatol. 2005;3(10 Suppl 2):S132–5.
Kamegaya Y, Hiasa Y, Zukerberg L, Fowler N, Blackard JT, Lin W, et al. Hepatitis C virus acts as a tumor accelerator by blocking apoptosis in a mouse model of hepatocarcinogenesis. Hepatology. 2005;41(3):660–7. https://doi.org/10.1002/hep.20621.
Koike K, Moriya K, Matsuura Y. Animal models for hepatitis C and related liver disease. Hepatol Res. 2010;40(1):69–82. https://doi.org/10.1111/j.1872-034X.2009.00593.x.
Schulze K, Imbeaud S, Letouze E, Alexandrov LB, Calderaro J, Rebouissou S, et al. Exome sequencing of hepatocellular carcinomas identifies new mutational signatures and potential therapeutic targets. Nat Genet. 2015;47(5):505–11. https://doi.org/10.1038/ng.3252.
Cancer Genome Atlas Research Network. Electronic address: wheeler@bcm.edu, Cancer Genome Atlas Research Network. Comprehensive and integrative genomic characterization of hepatocellular carcinoma. Cell. 2017;169(7):1327–41 e23. https://doi.org/10.1016/j.cell.2017.05.046.
Schulze K, Nault JC, Villanueva A. Genetic profiling of hepatocellular carcinoma using next-generation sequencing. J Hepatol. 2016;65(5):1031–42. https://doi.org/10.1016/j.jhep.2016.05.035.
Beroukhim R, Mermel CH, Porter D, Wei G, Raychaudhuri S, Donovan J, et al. The landscape of somatic copy-number alteration across human cancers. Nature. 2010;463(7283):899–905. https://doi.org/10.1038/nature08822.
Kawate S, Fukusato T, Ohwada S, Watanuki A, Morishita Y. Amplification of c-myc in hepatocellular carcinoma: correlation with clinicopathologic features, proliferative activity and p53 overexpression. Oncology. 1999;57(2):157–63. https://doi.org/10.1159/000012024.
Dang CV. MYC on the path to cancer. Cell. 2012;149(1):22–35. https://doi.org/10.1016/j.cell.2012.03.003.
Santoni-Rugiu E, Nagy P, Jensen MR, Factor VM, Thorgeirsson SS. Evolution of neoplastic development in the liver of transgenic mice co-expressing c-myc and transforming growth factor-alpha. Am J Pathol. 1996;149(2):407–28.
Shachaf CM, Kopelman AM, Arvanitis C, Karlsson A, Beer S, Mandl S, et al. MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancer. Nature. 2004;431(7012):1112–7. https://doi.org/10.1038/nature03043.
Calvisi DF, Conner EA, Ladu S, Lemmer ER, Factor VM, Thorgeirsson SS. Activation of the canonical Wnt/beta-catenin pathway confers growth advantages in c-Myc/E2F1 transgenic mouse model of liver cancer. J Hepatol. 2005;42(6):842–9. https://doi.org/10.1016/j.jhep.2005.01.029.
Zhan T, Rindtorff N, Boutros M. Wnt signaling in cancer. Oncogene. 2017;36(11):1461–73. https://doi.org/10.1038/onc.2016.304.
Khalaf AM, Fuentes D, Morshid AI, Burke MR, Kaseb AO, Hassan M, et al. Role of Wnt/beta-catenin signaling in hepatocellular carcinoma, pathogenesis, and clinical significance. J Hepatocell Carcinoma. 2018;5:61–73. https://doi.org/10.2147/JHC.S156701.
Nault JC, Mallet M, Pilati C, Calderaro J, Bioulac-Sage P, Laurent C, et al. High frequency of telomerase reverse-transcriptase promoter somatic mutations in hepatocellular carcinoma and preneoplastic lesions. Nat Commun. 2013;4:2218. https://doi.org/10.1038/ncomms3218.
Inagawa S, Itabashi M, Adachi S, Kawamoto T, Hori M, Shimazaki J, et al. Expression and prognostic roles of beta-catenin in hepatocellular carcinoma: correlation with tumor progression and postoperative survival. Clin Cancer Res. 2002;8(2):450–6.
Lai TY, Su CC, Kuo WW, Yeh YL, Kuo WH, Tsai FJ, et al. β-catenin plays a key role in metastasis of human hepatocellular carcinoma. Oncol Rep. 2011;26(2):415–22. https://doi.org/10.3892/or.2011.1323.
Hoshida Y, Nijman SM, Kobayashi M, Chan JA, Brunet JP, Chiang DY, et al. Integrative transcriptome analysis reveals common molecular subclasses of human hepatocellular carcinoma. Cancer Res. 2009;69(18):7385–92. https://doi.org/10.1158/0008-5472.CAN-09-1089.
Harada N, Miyoshi H, Murai N, Oshima H, Tamai Y, Oshima M, et al. Lack of tumorigenesis in the mouse liver after adenovirus-mediated expression of a dominant stable mutant of beta-catenin. Cancer Res. 2002;62(7):1971–7.
Harada N, Oshima H, Katoh M, Tamai Y, Oshima M, Taketo MM. Hepatocarcinogenesis in mice with beta-catenin and Ha-ras gene mutations. Cancer Res. 2004;64(1):48–54.
Nejak-Bowen KN, Thompson MD, Singh S, Bowen WC Jr, Dar MJ, Khillan J, et al. Accelerated liver regeneration and hepatocarcinogenesis in mice overexpressing serine-45 mutant beta-catenin. Hepatology. 2010;51(5):1603–13. https://doi.org/10.1002/hep.23538.
Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–74. https://doi.org/10.1016/j.cell.2011.02.013.
Kastenhuber ER, Lowe SW. Putting p53 in context. Cell. 2017;170(6):1062–78. https://doi.org/10.1016/j.cell.2017.08.028.
Hussain SP, Schwank J, Staib F, Wang XW, Harris CC. TP53 mutations and hepatocellular carcinoma: insights into the etiology and pathogenesis of liver cancer. Oncogene. 2007;26(15):2166–76. https://doi.org/10.1038/sj.onc.1210279.
Chawanthayatham S, Valentine CC 3rd, Fedeles BI, Fox EJ, Loeb LA, Levine SS, et al. Mutational spectra of aflatoxin B1 in vivo establish biomarkers of exposure for human hepatocellular carcinoma. Proc Natl Acad Sci U S A. 2017;114(15):E3101–E9. https://doi.org/10.1073/pnas.1700759114.
Lewis BC, Klimstra DS, Socci ND, Xu S, Koutcher JA, Varmus HE. The absence of p53 promotes metastasis in a novel somatic mouse model for hepatocellular carcinoma. Mol Cell Biol. 2005;25(4):1228–37. https://doi.org/10.1128/MCB.25.4.1228-1237.2005.
Farazi PA, Glickman J, Horner J, Depinho RA. Cooperative interactions of p53 mutation, telomere dysfunction, and chronic liver damage in hepatocellular carcinoma progression. Cancer Res. 2006;66(9):4766–73. https://doi.org/10.1158/0008-5472.CAN-05-4608.
Chen YW, Klimstra DS, Mongeau ME, Tatem JL, Boyartchuk V, Lewis BC. Loss of p53 and Ink4a/Arf cooperate in a cell autonomous fashion to induce metastasis of hepatocellular carcinoma cells. Cancer Res. 2007;67(16):7589–96. https://doi.org/10.1158/0008-5472.CAN-07-0381.
Cullen JM, Sandgren EP, Brinster RL, Maronpot RR. Histologic characterization of hepatic carcinogenesis in transgenic mice expressing SV40 T-antigens. Vet Pathol. 1993;30(2):111–8. https://doi.org/10.1177/030098589303000203.
Viatour P, Ehmer U, Saddic LA, Dorrell C, Andersen JB, Lin C, et al. Notch signaling inhibits hepatocellular carcinoma following inactivation of the RB pathway. J Exp Med. 2011;208(10):1963–76. https://doi.org/10.1084/jem.20110198.
Hopkins BD, Parsons RE. Molecular pathways: intercellular PTEN and the potential of PTEN restoration therapy. Clin Cancer Res. 2014;20(21):5379–83. https://doi.org/10.1158/1078-0432.CCR-13-2661.
Sun H, Lesche R, Li DM, Liliental J, Zhang H, Gao J, et al. PTEN modulates cell cycle progression and cell survival by regulating phosphatidylinositol 3,4,5,-trisphosphate and Akt/protein kinase B signaling pathway. Proc Natl Acad Sci U S A. 1999;96(11):6199–204.
Horie Y, Suzuki A, Kataoka E, Sasaki T, Hamada K, Sasaki J, et al. Hepatocyte-specific Pten deficiency results in steatohepatitis and hepatocellular carcinomas. J Clin Invest. 2004;113(12):1774–83. https://doi.org/10.1172/JCI20513.
Shen WH, Balajee AS, Wang J, Wu H, Eng C, Pandolfi PP, et al. Essential role for nuclear PTEN in maintaining chromosomal integrity. Cell. 2007;128(1):157–70. https://doi.org/10.1016/j.cell.2006.11.042.
Kotelevets L, van Hengel J, Bruyneel E, Mareel M, van Roy F, Chastre E. Implication of the MAGI-1b/PTEN signalosome in stabilization of adherens junctions and suppression of invasiveness. FASEB J. 2005;19(1):115–7. https://doi.org/10.1096/fj.04-1942fje.
Hu TH, Huang CC, Lin PR, Chang HW, Ger LP, Lin YW, et al. Expression and prognostic role of tumor suppressor gene PTEN/MMAC1/TEP1 in hepatocellular carcinoma. Cancer. 2003;97(8):1929–40. https://doi.org/10.1002/cncr.11266.
Di Cristofano A, Pesce B, Cordon-Cardo C, Pandolfi PP. Pten is essential for embryonic development and tumour suppression. Nat Genet. 1998;19(4):348–55. https://doi.org/10.1038/1235.
Stambolic V, Tsao MS, Macpherson D, Suzuki A, Chapman WB, Mak TW. High incidence of breast and endometrial neoplasia resembling human Cowden syndrome in pten+/− mice. Cancer Res. 2000;60(13):3605–11.
Podsypanina K, Ellenson LH, Nemes A, Gu J, Tamura M, Yamada KM, et al. Mutation of Pten/Mmac1 in mice causes neoplasia in multiple organ systems. Proc Natl Acad Sci U S A. 1999;96(4):1563–8.
Blackburn EH. Structure and function of telomeres. Nature. 1991;350(6319):569–73. https://doi.org/10.1038/350569a0.
Cong YS, Wright WE, Shay JW. Human telomerase and its regulation. Microbiol Mol Biol Rev. 2002;66(3):407–25, table of contents.
Park YM, Choi JY, Byun BH, Cho CH, Kim HS, Kim BS. Telomerase is strongly activated in hepatocellular carcinoma but not in chronic hepatitis and cirrhosis. Exp Mol Med. 1998;30(1):35–40. https://doi.org/10.1038/emm.1998.5.
Nagao K, Tomimatsu M, Endo H, Hisatomi H, Hikiji K. Telomerase reverse transcriptase mRNA expression and telomerase activity in hepatocellular carcinoma. J Gastroenterol. 1999;34(1):83–7.
Blasco MA, Lee HW, Hande MP, Samper E, Lansdorp PM, DePinho RA, et al. Telomere shortening and tumor formation by mouse cells lacking telomerase RNA. Cell. 1997;91(1):25–34.
Lechel A, Holstege H, Begus Y, Schienke A, Kamino K, Lehmann U, et al. Telomerase deletion limits progression of p53-mutant hepatocellular carcinoma with short telomeres in chronic liver disease. Gastroenterology. 2007;132(4):1465–75. https://doi.org/10.1053/j.gastro.2007.01.045.
Farazi PA, Glickman J, Jiang S, Yu A, Rudolph KL, DePinho RA. Differential impact of telomere dysfunction on initiation and progression of hepatocellular carcinoma. Cancer Res. 2003;63(16):5021–7.
Grivennikov SI, Greten FR, Karin M. Immunity, inflammation, and cancer. Cell. 2010;140(6):883–99. https://doi.org/10.1016/j.cell.2010.01.025.
Maeda S, Kamata H, Luo JL, Leffert H, Karin M. IKKbeta couples hepatocyte death to cytokine-driven compensatory proliferation that promotes chemical hepatocarcinogenesis. Cell. 2005;121(7):977–90. https://doi.org/10.1016/j.cell.2005.04.014.
Luedde T, Beraza N, Kotsikoris V, van Loo G, Nenci A, De Vos R, et al. Deletion of NEMO/IKKgamma in liver parenchymal cells causes steatohepatitis and hepatocellular carcinoma. Cancer Cell. 2007;11(2):119–32. https://doi.org/10.1016/j.ccr.2006.12.016.
He G, Karin M. NF-kappaB and STAT3 – key players in liver inflammation and cancer. Cell Res. 2011;21(1):159–68. https://doi.org/10.1038/cr.2010.183.
Pikarsky E, Porat RM, Stein I, Abramovitch R, Amit S, Kasem S, et al. NF-kappaB functions as a tumour promoter in inflammation-associated cancer. Nature. 2004;431(7007):461–6. https://doi.org/10.1038/nature02924.
Sanderson N, Factor V, Nagy P, Kopp J, Kondaiah P, Wakefield L, et al. Hepatic expression of mature transforming growth factor beta 1 in transgenic mice results in multiple tissue lesions. Proc Natl Acad Sci U S A. 1995;92(7):2572–6.
Newell P, Villanueva A, Friedman SL, Koike K, Llovet JM. Experimental models of hepatocellular carcinoma. J Hepatol. 2008;48(5):858–79. https://doi.org/10.1016/j.jhep.2008.01.008.
Gao GP, Alvira MR, Wang L, Calcedo R, Johnston J, Wilson JM. Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc Natl Acad Sci U S A. 2002;99(18):11854–9. https://doi.org/10.1073/pnas.182412299.
Nakai H, Fuess S, Storm TA, Muramatsu S, Nara Y, Kay MA. Unrestricted hepatocyte transduction with adeno-associated virus serotype 8 vectors in mice. J Virol. 2005;79(1):214–24. https://doi.org/10.1128/JVI.79.1.214-224.2005.
Liu F, Song Y, Liu D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther. 1999;6(7):1258–66. https://doi.org/10.1038/sj.gt.3300947.
Zhang G, Budker V, Wolff JA. High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA. Hum Gene Ther. 1999;10(10):1735–7. https://doi.org/10.1089/10430349950017734.
Kamimura K, Yokoo T, Abe H, Kobayashi Y, Ogawa K, Shinagawa Y, et al. Image-guided hydrodynamic gene delivery: current status and future directions. Pharmaceutics. 2015;7(3):213–23. https://doi.org/10.3390/pharmaceutics7030213.
Aronovich EL, McIvor RS, Hackett PB. The Sleeping Beauty transposon system: a non-viral vector for gene therapy. Hum Mol Genet. 2011;20(R1):R14–20. https://doi.org/10.1093/hmg/ddr140.
Wuestefeld T, Pesic M, Rudalska R, Dauch D, Longerich T, Kang TW, et al. A Direct in vivo RNAi screen identifies MKK4 as a key regulator of liver regeneration. Cell. 2013;153(2):389–401. https://doi.org/10.1016/j.cell.2013.03.026.
Xue W, Chen S, Yin H, Tammela T, Papagiannakopoulos T, Joshi NS, et al. CRISPR-mediated direct mutation of cancer genes in the mouse liver. Nature. 2014;514(7522):380–4. https://doi.org/10.1038/nature13589.
Chen X, Calvisi DF. Hydrodynamic transfection for generation of novel mouse models for liver cancer research. Am J Pathol. 2014;184(4):912–23. https://doi.org/10.1016/j.ajpath.2013.12.002.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Nature Switzerland AG
About this chapter
Cite this chapter
Molina-Sánchez, P., Lujambio, A. (2019). Experimental Models for Preclinical Research in Hepatocellular Carcinoma. In: Hoshida, Y. (eds) Hepatocellular Carcinoma. Molecular and Translational Medicine. Humana, Cham. https://doi.org/10.1007/978-3-030-21540-8_16
Download citation
DOI: https://doi.org/10.1007/978-3-030-21540-8_16
Published:
Publisher Name: Humana, Cham
Print ISBN: 978-3-030-21539-2
Online ISBN: 978-3-030-21540-8
eBook Packages: MedicineMedicine (R0)