Hepatocellular carcinoma mouse models: Hepatitis B virus-associated hepatocarcinogenesis and haploinsufficient tumor suppressor genes

Table 1 Hepatitis B virus-related transgenic mice

Transgene	Promoter	Expression	Pathology	Ref.
PreS/S/X	Endogenous	Surface proteins	Until 6 mo: no obvious pathology	[28]
PreS/S/X (two TGs)	Endogenous and mouse metallothionein I	Surface proteins	Not determined	[29]
PreS/S/X	Mouse metallothionein I and Albumin	Surface proteins	Ground glass	[22,30-32,34]
			7-9 mo: adenoma
			12 mo: HCC
			18 mo: HCC (100%)
Genome (2X)	Endogenous	Surface proteins	Not determined	[241]
PreS/S (Knock-in)	Mouse p21	Surface proteins	15-24 mo: HCC [53% (♂KI/+), 72% (♂KI/KI), 0% (♀KI/+ and KI/KI)]	[37]
PreS/S	Endogenous	Surface proteins	HCC	[33,40]
X	Human α-1-antitrypsin	HBx protein	Focal necrosis, hyperplasia nodule (Authors claimed that X is not tumorigenic)	[42]
X	Endogenous	HBx protein	4 mo: altered hepatocyte in multifocal area	[43,44]
			10 mo: tumor nodule
			16 mo: tumor (80%)
			24 mo: HCC (84%)
X	Endogenous	HBx protein	6 mo: neoplastic nodules (66.6%)	[41]
			15-18 mo: small neoplastic nodule (100%) and HCC (75%)
X (Knock-in)	Mouse p21	HBx protein	15-24 mo: HCC [60%-64% (♂KI/+ and KI/KI), 43%-46% (♀KI/+ and KI/KI)]	[37]
X	Mouse albumin	HBx protein	12 mo: dysplasia nodule (100%)	[45-47]
			16 mo: HCC (80%)
			20 mo: HCC (90%-100%)
Precore/core	Mouse metallothionein I	Precore protein	Not determined	[56]
Precore/core	Endogenous	Core protein	12 mo: no hepatitis or HCC	[13,57]
Precore/core (two TGs)	Endogenous	Precore and core proteins	Not determined	[58]
Genome (2X)	Endogenous	Replicative intermediates	10 mo: no obvious pathology	[60]
Genome (1.2X)	Endogenous	Replicative virus	12 mo: no hepatitis or HCC	[59,61]
			24 mo: no obvious pathology
Genome (1.3X)	Endogenous	Replicative intermediates	Until 12 mo: no obvious pathology	[62]

PreS/S: Coding region of hepatitis B virus (HBV) large, middle and small surface proteins; X: Coding region of HBV X protein; Precore/core: Coding region of HBV pre-core/core proteins; KI: Knock-in allele; TG: Transgenic mice; Genome (2X): Indicates 2 copies of the complete 3.2-kilobase HBV genome.

Table 2 Mouse models of tumor suppressor genes in hepatocellular carcinoma

Official gene name (common name)	Mouse genotype	Mouse genetic background	Pathology	TGS type	Functions	Human Chromosome location	Expression in human HCC tumor	Ref.
Autophagy regulator
Becn1 (Beclin1)	Beclin1+/-	Mixed (129Sv/J; C57BL/6J)	16-18 mo: HCC (8%)	Haplo-insufficiency	Regulation of autophagy	17q21	Down	[85-87]
			18-22 mo: HCC (26%)
			Promote HBV-mediated HCC
Atg5	Alb-Cre; Atg5flx/flx	C57BL/6	2 mo: hepatic cell death, proliferation, inflammation, fibrosis and hepatomegaly	Recessive	Control autophagosome formation	6q21	Down	[90,91]
			12 mo: hepatic adenoma (100%)
Atg7	Alb-Cre; Atg7flx/flx;	C57BL/6	4 mo: fatty liver	Recessive	Control autophagosome formation	3p25.3	Up	[92-94]
			12 mo: hepatic adenoma (90.9%)
Cell cycle regulator
Klf6	KLF6+/-	C57BL/6	Promote DEN-induced HCC	Haplo-insufficiency	Cell cycle regulation	10p15	Down	[99,101]
Plk4	Plk4+/-	Mixed (129Sv;CD1)	3 mo: mitotic failure in hepatocyte regeneration	Haplo-insufficiency	Control of cell cycle progression	4q28	Down	[104,105]
			18-24 mo: HCC (30%)
Cdkn1a (p21)	p21+/-	Mixed (129Sv; C57BL/6)	Promote γ-irradiation-induced HCC	Haplo-insufficiency	Inhibits the activity of CDK2 or CDK4	6p21.2	Up	[107,242]
	p21-/-	C57BL/6	Promote HCC in Nemo liver-specific KO	Recessive				[108]
Trp53 (p53)	p53+/-	C57BL/6	Promote HBV-mediated and AFB1-induced HCC	Haplo-insufficiency	DNA damage response transcription factor	17p13.1	Up	[111,243]
	Aflp-Cre; p53flx/flx	C57BL/6	14-20 mo: HCC (90%)	Recessive				[112]
Protein ubiquination
Amfr (Gp78)	Gp78-/-	C57BL/6	12 mo: fatty liver, inflammation and HCC (24%)	Recessive	E3 ubiquitin ligase	16q21	Down	[117]
Park2 (Parkin)	Parkin-/-	Mixed (129/C57BL6SJL)	11-12 mo: hepatocyte proliferation and hepatomegaly	Recessive	E3 ubiquitin ligase	6q25.2-q27	Down	[118,122]
			16-17 mo: HCC (33%)
			22-23 mo: HCC (45%)
Trim24	Trim24-/-	C57BL/6	2 mo: hepatic cell death and inflammation	Recessive	E3 ubiquitin ligase	7q32	Up	[119,125]
			4-6 mo: fatty liver, fibrosis and liver nodule
			9-21 mo: HCC (55%)
Genome stability maintenance
Anxa7 (Annexin 7)	Anx7+/-	C57BL/6	10 mo: Hepatomegaly	Haploinsufficiency	Ca2+-dependent endocrine secretion	10q22.2	Up	[131,132]
			12 mo: HCC (3.6%)
Nbn (Nibrin)	Nbn+/-	Mixed (129Sv; C57BL/6)	23 mo: HCC (8.6%)	Haploinsufficiency	DNA repair	8q21	Up	[134,244]
Pinx1	PinX1+/-	Mixed (129Sv; C57BL/6)	9-18 mo: HCC (17.7%)	Haploinsufficiency	Telomerase inhibitor	8p23	Down	[136,137]
Sgol1 (Shugoshin 1)	Sgo1+/-	C57BL/6	4 mo: hepatocyte DNA damage	Haploinsufficiency	Protector of chromosome cohesion and centrosome integrity	3p24.3	Up	[139,140]
			12 mo: HCC
			Promote AOM-induced HCC
Metabolic function
Acox1 (Acyl-CoA oxidase 1)	Aox-/-	Mixed (129/Ola; C57BL/6)	2-4 mo: fatty liver, inflammation and hepatocyte proliferation	Recessive	Peroxisomal β-oxidation	17q25.1	Not determined	[144]
			10-15 mo: HCC (100%)
Bhmt	Bhmt-/-	C57BL/6	1 mo: fatty liver	Recessive	Methionine metabolism	5q14.1	Down	[149,150]
			12 mo: HCC (50%)
Nr1h4 (Farnesoid X receptor)	Fxr-/-	C57BL/6N	3 mo: hepatic cell death and proliferation	Recessive	Transcriptional regulation of synthesis and transport of bile acids	12q23.1	Down	[155,157]
			6-9 mo: fatty liver and inflammation
			12 mo: fibrosis and HCC (16.1%)
Gnmt (Glycine N-methyltransferase)	Gnmt-/-	Mixed (129Sv; C57BL/6)	3 mo: fatty liver and fibrosis	Recessive	Methionine metabolism	6p12	Down	[148,151]
			8 mo: HCC (100%) at 8-mo
Abcb4 (Mdr2)	Mdr2-/-	129/OlaHsd	3 mo: inflammation and fibrosis	Recessive	Phosphatidylcholine translocase	7q21.1	Down	[156,158]
			6-mo: HCC
Hippo signaling pathway
Stk4 (Mst1)	Alb-Cre; Mst1/2flx/flx	Mixed (129Sv; C57BL/6;CD1)	1 mo: hepatocyte proliferation and hepatomegaly	Recessive	Kinase (negative regulator)	20q11.2	Not determined	[165,166]
			6 mo: HCC (100%)
Stk3 (Mst2)	Alb-Cre; Mst1/2flx/flx	Mixed (129Sv; C57BL/6;CD1)	1 mo: hepatocyte proliferation and hepatomegaly	Recessive	Kinase (negative regulator)	8q22.2	Not determined	[165,166]
			6 mo: HCC (100%)
Nf2 (Neurofibromin 2)	Alb-Cre; Nf2flx/flx	Mixed (FVB/N; C57BL/6)	2 mo: oval cell over-proliferation and hepatomegaly	Recessive	Negative regulator	22q12.2	Not determined	[168]
			7 mo: HCC (100%)
Jak/Stat signaling pathway
Socs1	Socs1+/-	C57BL/6	Promote DEN-induced HCC	Haploinsufficiency	Negative regulator	16p13.13	Down	[174]
Socs3	Socs3+/-	Mixed (129Sv; C57BL/6)	Promote DEN-induced HCC	Haploinsufficiency	Negative regulator	17q25.3	Down	[176]
Ptpn11 (Shp2)	Alb-Cre; Shp2flx/flx	C57BL/6	2-3 mo: inflammation and hepatic cell death	Recessive	Protein tyrosine phosphatase (negative regulator)	12q24	Down	[179]
			8-mo: hyperplasia nodule
			12-18 mo: hepatic adenoma (68%)
			Promote DEN-induced HCC
Ptpro	Ptpro-/-	C57BL/6	Promote DEN-induced HCC	Recessive	Receptor-like protein tyrosine phosphatase (negative regulator)	12p13.3	Down	[181,182]
NF-κB signaling pathway
Cyld (Cylindromatosis)	Alfp-Cre; Cyldflx/flx	C57BL/6 congenic	1-2 mo: hepatic cell death, inflammation and fibrosis	Recessive	Deubiquitinase (negative regulator)	16q12.1	Down	[188,189]
			12 mo: HCC
Lgals3 (Galectin-3)	Gal3-/-	CD1	6 mo: fatty liver and inflammation	Recessive	Regulation of inflammatory responses	14q22.3	Up	[192-194]
			15 mo: fibrosis and liver nodule
			25 mo: HCC (100%)
Ikbkg (Nemo)	Alfp-Cre; Nemoflx/flx	C57BL/6	2 mo: fatty liver, inflammation, hepatic cell death and proliferation	Recessive	Activation of NF-κB	Xq28	Down	[195,196]
			6 mo: dysplastic nodule
			12 mo: HCC (100%)
Map3k7 (Tak1)	Alfp-Cre; Tak1flx/flx	Mixed (129/Ola; C57BL/6)	1-2 mo: hepatic cell death, proliferation and fibrosis	Recessive	Serine/threonine protein kinase (Activation of the NF-κB)	6q15	Not determined	[197]
			4-8 mo: HCC (88%)
PI3K/Akt/mTOR signaling pathway
Stk11 (Lkb1)	Lkb1+/-	Mixed (129Sv; C57BL/6)	10-12 mo: Hepatic hyperplasia, HCC (29%)	Recessive	Serine/threonine kinase (activator of AMPK)	19p13.3	Down	[201,202]
			14 mo: HCC (75%)
Pten	Alb-Cre; Ptenflx/flx	C57BL/6	2-3 mo: fatty liver and hepatomegaly	Recessive	Phospholipid phosphatase (negative regulator)	10q23.3	Down	[204,208]
			9-10 mo: inflammation and fibrosis
			17-18 mo: HCC (66%)
Raptor (Regulatory associated protein of mTOR, complex 1)	Alb-Cre; Raptorflx/flx	C57BL/6	2 mo: hepatic cell death, inflammation and fibrosis	Recessive	Activation of mTOR activity	17q25.3	Up	[209,210]
			Promote DEN-induced HCC
TGF-β signaling pathway
Sptbn1 (β-spectrin)	Elf+/-	Mixed (129SvEv; Black Swiss)	15 mo: Fatty liver, HCC (40%)	Haploinsufficiency	Propagation of TGF-β signal	2p21	Down	[214]
Tgfb1 (TGF-β1)	Tgf-β1+/-	C57BL/6NCr	Promote DEN-induced HCC	Haploinsufficiency	Growth factor	19q13.1	Up	[216,245]
Tgfbr2 (TGF-β type II receptor)	TβR-II+/-	Mixed (129Sv; C57BL/6)	Promote DEN-induced HCC	Recessive	TGF-β receptor	3p22	Down	[217,218]
Wnt signaling pathway
Apc	Adenovirus-Cre; Apcflx/flx	C57BL/6N	9 mo: HCC (67%)	Recessive	Antagonist	5q21	Down	[222,223]
MicroRNA
Mir122	Mir122a-/-	C57BL/6	3 mo: fatty liver, inflammation and fibrosis	Recessive	Post-transcriptional regulation of gene expression	18q21.31	Down	[227,228]
			11 mo: HCC (75%)
Mir140	Mir140-/-	C57BL/6	Promote DEN-induced HCC	Recessive	Post-transcriptional regulation of gene expression	16q22.1	Not determined	[229]
Miscellaneous
Ncoa5 (Nuclear receptor coactivator 5)	Ncoa5+/-	Mixed (129Sv; C57BL/6)	6 mo: fatty liver	Haploinsufficiency	Estrogen receptor coactivator	20q13.12	Down	[231]
			10 mo: inflammation and fibrosis
			10-18 mo: HCC (94%)
Prkar1a	Prkar1a+/-	Mixed (129Sv/J; C57BL/6)	9-19 mo: HCC (29.4%)	Haploinsufficiency	Regulation of the serine/ threonine kinase activity	17q24.2	Up	[233,246]
Ncoa2 (Nuclear receptor coactivator 2)	Ncoa2-/-	Mixed (129Sv/J; C57BL/6)	Promote DEN-induced HCC	Recessive	Transcriptional coactivator	8q13.3	Down	[235]
Nfe2l1 (Nuclear factor, erythroid 2-like 1)	Alb-Cre; Nrf1flx/flx	Mixed (129Sv; C57BL/6)	1-2 mo: fatty liver, necrosis and inflammation	Recessive	Transcription factor	17q21.3	Not determined	[238]
			6 mo: fibrosis
			12 mo: HCC (100%)

HCC: Hepatocellular carcinoma.