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. 2018 Feb 28;2018:6328970. doi: 10.1155/2018/6328970

Table 1.

Description of the main characteristics of the studies using flavonoids in the treatment of hepatocellular carcinoma in murine models.

Ref. Country Article Plant family/species Used part of plant Flavonoids type Extraction and purification method In vitro assay Animal model/strain Sex Age/weight Administration/
frequency
Induction of carcinoma concentration/
volume
Tumor measurement/
frequency
In vivo analysis
Wan et al., 2014 [15] China Preparation of morusin from Ramulus mori and its effects on mice with transplanted H22 hepatocarcinoma Moraceae/Ramulus mori Root bark Isoprenylated flavone (morusin) Ethanol extraction/Sephadex gel LH-20 and RF-HPLC ? Mice/SPF ICR 3–5 weeks/18–20 g Intraperitoneal/
daily for 10 days
Transplanted H22 cells
1 × 107 cells/ml/0.1 ml
Weight of tumor/2 weeks H&E, qRT-PCR (P53, survivin, cyclin D1, caspase-3, NF-Kβ)

Shu et al., 2014 [16] China Kurarinol induces hepatocellular carcinoma cell apoptosis through suppressing cellular signal transducer and activator of transcription 3 signaling Leguminosae/
Sophora flavescens
Root Kushenol H, kurarinol, norkurarinol, kushenol N, kurarinone Ethanol extraction, HPLC and Sephadex LH-20 column HepG2, Huh-7, Bel-7402 cells humans, HL-7702, H22 cells mouse Mice/Kunming ?/18–22 g Intraperitoneal/
daily for 10 days
Transplant of H22 cells
1 × 106 cells/ml/?
Weight of tumor/experiment end Serum (white blood, red blood cells, platelets, ALT, AST, BUN, uric acid, and CK), TUNEL, immunohistochemistry (STAT3)

Wang et al., 2014 [17] China Liquiritigenin induces tumor cell death through mitogen-activated protein kinase- (MPAKs-) mediated pathway in hepatocellular carcinoma cells ?/Glycyrrhiza radix ? Liquiritigenin (LQ) Purchased PLC/PRF/5, HepG2 human cells Mice/BALB/cA nude 5 weeks/? Intraperitoneal/
daily for 18 days
Transplanted PLC/PRF/5
5 × 107 cells/ml/0.1 ml
Volume and diameters of tumor (mm3), body weight/every day ?

Zhang et al., 2014 [18] China Dihydromyricetin promotes hepatocellular carcinoma regression via a p53 activation-dependent mechanism ?/Hovenia dulcis ? Dihydromyricetin (DHM) Purchased HepG2, SMMC-7721, HL7702, L02 cells, primary cell: 4401, 4403, 1204 Mice/BALB/cA nude 8–10 weeks/? ?/daily for 21 days Transplanted HepG2 cells
1 × 107 cells/ml/0.2 ml
? Immunohistochemistry (P53)

Darweish et al., 2014 [19] Egypt Chemopreventive and hepatoprotective effects of epigallocatechin gallate against hepatocellular carcinoma: role of heparan sulfate proteoglycans pathway ?/Camellia sinensis ? Epigallocatechin gallate (EGCG) ? HepG2 cell Rats/Sprague Dawley ?/180–200 g ?/twice for week for 16 days Thioacetamide (200 mg/Kg b.w.) ? H&E, immunohistochemistry (HSPGs), qRT-PCR (FGF-2), in serum (AST, albumin, and bilirubin), enzymatic determination (MDA, SOD, MPO, ELISA MMP-9, HSPGs, AFP, and syndecan-1)

Zheng et al., 2014 [20] China/USA Anticancer effects of Baicalein on hepatocellular carcinoma cells ?/Scutellaria sp. Radix Baicalein Purchased MIHAs, H22, BEL-7404, HepG2 cells Mice/ICR ?/18–22 g Intraperitoneal/
daily for 12 days
Transplanted H22
2 × 105 cells/ml/0.2 ml
Weight of tumors/experiment end Immunohistochemistry (AKT, p-AKT Ser473, β-catenin, cyclin D1)

Fan et al., 2014 [21] China Luteoloside suppresses proliferation and metastasis of hepatocellular carcinoma cells by Inhibition of NLRP3 inflammasome ?/Gentiana macrophylla ? Luteoloside Purchased Hep3B, SNU-449, Huh-7, SMMC-7721, MHCCLM3, MHCC97-H cell Mice/BALB/cA nude 6 weeks/? Oral/daily for 4 weeks (subcutaneous) and 8 weeks (metastasis) groups Transplanted SMMC-7721
2 × 106 cells 7/0.2 ml
Volume of tumor (cm3)/every 3 to 4 days H&E

Feng et al., 2014 [22] China Effect of grape procyanidins on tumor angiogenesis in liver cancer xenograft models ? Seed Proanthocyanidins (GPC) ? ? Mice/ SPF Kunming ? 4–6 weeks/20 g Intraperitoneal/
daily for 10 days
Transplanted H22 cells
1 × 106 cell/ml/0.2 ml
? Immunohistochemistry (VEGF and CD34), RT-PCR (VEGF and β-actin)

Zhang et al., 2013 [23] China Silybin-mediated inhibition of notch ?/Silybum marianum ? Silybin (SIL) ? HepG2 cells Mice/athymic nude 4–6 weeks/? Intraperitoneal/5 times for week for 20 days Transplanted HepG2
1 × 106 cells/ml/?
Size of tumor/every 3 days Western blot (Notch1, Hes1, RBP-Jκ, survivin, cyclin D1, Bcl2, BAX, β-actin)

Yang et al., 2013 [24] China Antitumor effects of two extracts from Oxytropis falcata on hepatocellular carcinoma in vitro and in vivo ?/Oxytropis falcata Whole plant Flavonoids (FOF) 2′,4′-dihydroxychalcone Ethanol extraction/? SMMC-7721 cells Mice/ICR 5-6 weeks/18–22 g Intragastric/? Transplanted H22 cells
5 × 106 cell/ml/0.2 ml
Tumor growth inhibitory ratio/experiment end ?

Yu et al., 2013 [25] China A study on the antitumor effect of total flavonoids from Pteris multifida poir in H22 tumor-bearing mice ?/Pteris multifida Whole plant Total flavonoids Ethanol extraction/? ? Mice/Kunming 1/2 ♂ and 1/2 ♀ ?/18–22 g Intraperitoneal/
daily for 10 days
Transplanted H22 cell
5 × 107 cells/ml/0.3 ml
Tumor inhibition rate and organ index (spleen and thymus)/experiment end ELISA (TNF-α and IL-2), enzymatic determination (T-AOC, MDA)

Xiang et al., 2013 [26] China Chemical composition of total flavonoids from Salvia chinensia Benth and their proapoptotic effect on hepatocellular carcinoma cells: potential roles of suppressing cellular NF-kB signaling ?/Salvia chinensia Whole plant Total flavonoids Alcoholic extraction 95%/HPLC and chromatography in silica gel column HepG2, Huh-7 HCC cells Mice/Kunming ?/18–22 g Intraperitoneal/
daily for 10 days
Transplanted H22 cells
1 × 106 cells/ml/?
Weight of tumor, organ index/experiment end Automatic counting (red blood cells, hemoglobin, white blood cells, platelets), serum (AST, ALT, BUN, CK, uric acid), ELISA (caspase-3, caspase-8, caspase-9)

Hashimoto et al., 2014 [27] Japan Methylated-(3′′)-epigallocatechin gallate analog suppresses tumor growth in Huh7 hepatoma cells via inhibition of angiogenesis ?/Camellia sinensis Green leaves Polyphenolic, catechin, methylated-(3)-epigallocatechin gallate (MethylEGCG) Purchased/HPLC purification HUVECs, Huh7 cells Mice/BALB/cA nude 6 weeks/20 g Intraperitoneal/
daily for 21 days
Transplanted Huh7 cells
5 × 106 cell/ml/?
Size, volume of tumor/every week Immunohistochemistry (CD31)

Saleem et al., 2013 [28] India and Saudi Arabia Anticancer potential of rhamnocitrin 40-b-D-galactopyranoside against N-diethylnitrosamine-induced hepatocellular carcinoma in rats ?/Astragalus hamosus Leaves Flavonol glycoside (rhamnocitrin 40-b-D-galactopyranoside RGP) MeOH/H2O extraction/column Sephadex LH-20 ? Rats/Wistar ? ?/150–220 g Intraperitoneal/
daily for 20 weeks
Single intraperitoneal dose of N-nitrosodiethylamine (NDEA) (200 mg/kg b.w.) ? Serum (ALT, AST, ALP, total bilirubin, protein content, LPO), enzymatic determination (SOD, CAT, GPx, GST), H&E

Monga et al., 2013 [29] India Growth inhibition and apoptosis induction by (+)-Cyanidan-3-ol in hepatocellular carcinoma Fabaceae/Acacia catechu Heart wood Cyanidan-3-ol (CD-3) Ethanol extraction/HPLC HepG2 cells Mice/BALB/cA 6-7 weeks/26-27 g Oral/daily for 20 weeks Single intraperitoneal dose of NDEA (200 mg/kg b.w.) and carbon tetrachloride (CCl4, 3 ml/kg b.w.) thrice a week for six weeks Relative liver weight/experiment end Serum (AST, ALT, ALP, γ-GT, TSA, LASA), enzymatic determination (MDA, SOD, CAT, GPx, GR, GST, T-SH, GSH, PrPr-SHs), H&E, immunohistochemistry (p53, p65, c-jun)

Zhao et al., 2013 [30] China Enhanced 5-fluorouracil cytotoxicity in high COX-2 expressing hepatocellular carcinoma cells by Wogonin via the PI3K/AKT pathway ?/Scutellaria radix ? Wogonin 5,7-dihydroxy-8-methoxyflavone (WOG) ? HepG2, SMMC-7721 cells Mice/BALB/cA nude 35–40 days/18–22 g Intravenous/once every two days for 10 times Transplanted SMMC-7721 cells
1 × 106 cells/mouse/?
Tumor inhibitory ratio, weight, size, volume of tumor/experiment end ?

Wei et al., 2012 [31] China Antitumor and antiangiogenic effects of Macrothelypteris viridifrons and its constituents by HPLC-DAD/MS analysis ?/Macrothelypteris viridifrons Air dried roots 2-(1,4-Dihydroxy-cyclohexyl)-5,7-dihydroxy-chromone-4′-O-glucoside; protoapigenin-4′-O-glucoside; 2-(1,2-dihydroxy-4-oxo-cyclohex-5-enyl)-5,7-dihydroxy-chromone; 5,7,2′,5′-tetrahydroxy-flavanone-2′-O-glucoside; protoapigenin; 2-(1,4-dihydroxy-cyclohexyl)-5,7-dihydroxy-chromone; 5,7,2′,5′-tetrahydroxy-flavanone-2′-O-6′′-O-acetylglucoside; 5,6-dihydro-6-methoxyprotoapigenone; quercetin-3-O-rutinose; protoapigenone; apigenin-4-O-glucoside; kaempferol-3-O-rutinose; kaempferol-3,7-di-O-rhamnoside; kaempferol-3-O-rhamnoside; apigenin-4-O-rhamnoside ?/HPLC-DAD/MS Amethyst C18-P column HUVECs, H-22 cells Mice/Kunming ?/18–22 g Intraperitoneal/
daily for 10 days
Transplanted H22 cells
2 × 106 cells/ml/0.2 ml
Weight of tumor, tumor inhibitory ratio/experiment end Serum (red blood cell, white blood cell, platelet and hemoglobin), BUN, CRE, AST and ALT, TSA, LASA, enzymatic determination (MDA, SOD, CAT, GPx, GR, GST, T-SH, GSH, PrPr-SHs), H&E, immunohistochemistry (CD34, VEGF)

Wu et al., 2011 [32] Taiwan Suppression of hepatitis B virus × protein-mediated tumorigenic effects by ursolic acid Rubiaceae/Morinda citrifolia Leaves Ursolic acid and silymarin ? Huh7, HepG2, Hep3B cells Mice/nude ? 3 weeks/? Intraperitoneal/?/8 weeks Transplanted 6 2.2.15 cells
1 × 106 cell/ml/0.1 ml
Diameters of tumor/experiment end Serum (ALT, AST, BUN, CRE)

Wang et al., 2011 [33] Taiwan Solanum nigrum L. polyphenolic extract inhibits hepatocarcinoma cell growth by inducing G2/M phase arrest and apoptosis ?/Solanum nigrum Whole plant Total flavonoids Water extract (SNWE)/? HepG2 cells Mice/Athymic nude ? ? Intraperitoneal/?/35 days Transplanted HepG2 cells
5 × 106 cell/ml/0.4 ml
Volume of tumor/every week, final volume, wet weight of tumor/experiment end ?

Cai et al., 2011 [34] China Apigenin inhibits hepatoma cell growth through alteration of gene expression patterns ? ? Apigenin ? Huh7 cells Mice/BALB/c nude 5 weeks/16–18 g Intraperitoneal/?/30 days Transplanted Huh7 cells
2 × 106 cell/ml/?
Number of tumors, diameters, wet weight of tumor/experiment end H&E

Huang et al., 2010 [35] China Carbonyl reductase 1 as a novel target of (−)-epigallocatechin gallate against hepatocellular carcinoma ?/Camellia sinensis ? Epigallocatechin gallate (EGCG) ? Hep G2, Hep 3B, SMMC-7721 cells Mice/? ? ? Injected/?/15 days Transplanted SMMC-7721 cells and Hep3B/?/? Tumor growth, body weight/? Serum (ALT, AST, LDH, CK-MBB, LDH, MDA, cTnT)

Liang et al., 2010 [36] China and USA Green tea catechins augment the antitumor activity of doxorubicin in an in vivo mouse model for chemoresistant liver cancer ?/Camellia sinensis ? Catechin and epigallocatechin-3-gallate ? BEL-7404, BEL-7404/DOX cells Mice/ BALB/cA nude ♂ and ?? 4-5 weeks/13–17 g Intraperitoneal/?/33 days Transplanted BEL-7404/DOX HCC cells
5 × 107 cell/ml/0.2 ml
Dimensions of tumor, volume of tumor/every 2 days, tumor growth, body weight/experiment end Fluorospectrophotometry (DOX), immunohistochemistry, RT-PCR (MDR1)

Zhou et al., 2010 [37] China Inhibition of hepatoma 22 tumor by liquiritigenin ?/Glycyrrhiza glabra ? Liquiritigenin (LQ) Purchased ? Mice/ICR ?/20–22 g Intragastric/?/15 days Transplanted H22 cells
10 × 106 cell/ml/0.2 ml
Size, volume of tumor, body weight/every 3 days, Inhibition ratio of tumor growth, weight of tumor, organ index/experiment end H&E, electron microscopy (tumor ultrastructure), enzymatic determination (MDA)

Yang et al., 2009 [38] China Antiproliferative efficacy of icariin on HepG2 hepatoma and its possible mechanism of action ?/Herba epimedii ? Icariin (5-hydroxy 4-methoxy 8-isopentenyl 3-O-α-rhamnosyl 7-O-β-glucosyl flavone) Purchased HepG2 cells Mice/NMRI nude ? Oral/28 weeks Transplanted HepG2 cells
1 × 107 cell/ml/?
Volume of tumor, tumor growth inhibition/twice per week Immunohistochemistry (CD4, CD8, CD19)

Zhao et al., 2010 [39] China Synergistic effect of 5-fluorouracil and the flavonoid Oroxylin A on HepG2 human hepatocellular carcinoma and on H22 transplanted mice ?/Scutellariae Radix ? Oroxylin A ? HepG2 cells Mice/Kunming ?/18–22 g Oral/?/7 days Transplanted H22 cells
5 × 106 cells/ml/0.9 ml
Volume of tumor/experiment end ?

Cui et al., 2009 [40] USA Effects and mechanisms of Silibinin on human hepatocellular carcinoma xenografts in nude mice ?/Silybum marianum ? Silibinin ? HuH7 cells Mice/nude ? ? Oral/?/5 weeks Transplanted HuH7 cells
5 × 106 cell/ml/0.25 ml
Volumes of tumor/every week, weight of tumor/experiment end Immunoprecipitation (AFP, PTEN, binding interaction between p21, p27 with CDK4, binding of DP1 with E2F1), Western blotting (Ki-67, p21, p27, E2F1, CDK4, p-Rb, caspase-3, caspase-9, PTEN, AC-H3, AC-H4, p-AKT, p-survivin and p-ERK, Plk1, Chk1, SOD1), ELISA (NF-kB)

Umarani et al., 2008 [41] India Protective effect of Kalpaamruthaa in combating the oxidative stress posed by aflatoxin B1-induced hepatocellular carcinoma with special reference to flavonoid structure-activity relationship Anacardiaceae/
Semecarpus anacardium
Fruit Flavonoids Dried and powdered fruit/? ? Rats/? 8–10 weeks/120–150 g Oral/?/28 days Intraperitoneal dose of AFB1 (2 mg/kg b.w.) ? Serum (protein, LPO, lipid peroxides, G6PD, vitamin E, vitamin C, uric acid), enzymatic determination (SOD, CAT, GPx, GR)

Miura et al., 2007 [42] Japan Effect of apple polyphenol extract on hepatoma proliferation and invasion in culture and on tumor growth, metastasis, abnormal lipoprotein profiles in hepatoma-bearing rats ?/Malusdomestica Unripe apples Apple polyphenol extract (APE) Purchased AH109A cells Rats/Donryu 5 weeks/? Oral/?/21 days Transplanted AH109A cells
1 × 107 cells/ml/0.5 ml
Size of tumor/every day Serum (T-Ch, HDL-Ch), LPO, fecal steroid, AI

Selvendiran et al., 2006 [43] Japan Luteolin promotes degradation in signal transducer and activator of transcription 3 in human hepatoma cells: an implication for the antitumor potential of flavonoids ? Seeds Luteolin ?/HPLC HepG2, HLF, HAK-1B, IMR-32 cells Mice/BALB/cA nude 5 weeks/? Oral/?/6 weeks Transplanted HAK-1B cells
1 × 107 cell/ml/?
Size, volume (mm3) of tumor/weekly Immunoblotting (Tyr705-P-STAT3)

Nishikawa et al., 2006 [44] Japan A green tea polyphenol, epigallocatechin-3-gallate, induces apoptosis of human hepatocellular carcinoma, possibly through inhibition of Bcl-2 family proteins ?/Camellia sinensis ? Epigallocatechin-3-gallate ? HLE, HepG2, HuH-7, PLC/PRF/5 cells Mice/BALB/cA nude 28 weeks/? Oral/?/25 days Transplanted HLE cells
1 × 106 cell/ml/?
Volume of tumor/daily TUNEL, immunohistochemistry (Bcl-2a, Bcl-xl)

Premalatha and Sachdanandam, 1999 [45] India Semecarpus anacardium L. nut extract administration induces the in vivo antioxidant defense system in aflatoxin B1 mediated hepatocellular carcinoma ?/Semecarpus anacardium L Nuts Total flavonoids Purchased ? Rat/Wistar ?/100 g Oral/?/14 days Single intraperitoneal dose of AFB1 (2 mg/kg b.w.) Weight of liver and kidney/experiment end Total protein, enzymatic determination (GSH, uric acid, vitamin E, vitamin C, CYP, T-SH, NPSH)

Nishida et al., 1994 [46] Japan Inhibitory effects of (−)-epigallocatechin gallate on spontaneous hepatoma in C3H/HeNCrj mice and human hepatoma-derived PLC/PRF/5 cells ?/Camellia sinensis ? Epigallocatechin gallate (EGCG) ? PLC/PRF5 cells Mice/C3H/HenCrj 8 weeks/? Oral/?/65 weeks Spontaneous hepatocarcinogenesis # of tumor, diameter of liver/experiment end H&E, albumin, bilirubin, GPT, γ-GT, total cholesterol

Zhang et al., 2002 [47] Japan Effects of dietary powdered green tea and theanine on tumor growth and endogenous hyperlipidemia in hepatoma-bearing rats ?/Camellia sinensis ? ? ? ? Rat/Donryu 4 weeks/? Oral/?/14 days Transplanted AH109A cells
5 × 105 cell/ml/0.5 ml
Size, volume, diameter of tumor, radius tumor/every day, weight of tumor/experiment end Precipitation method (HDL, LDL, VLDL), enzymatic determination (T-Ch and HDL-ChM, TBARS), fecal extraction (neutral sterol, bile acid)

Klaunig, 1992 [48] USA Chemopreventive effects of green tea components on hepatic carcinogenesis ?/Camellia sinensis ? Epicatechin, epicatechin gallate, epigallocatechin, epigallocatechin gallate Methanol extraction/steel column with silicic acid Primary cells mouse hepatocytes Rat/B6C3F1 ? Oral/?/28 weeks Single intraperitoneal dose NDEA (90 mg/kg b.w.) ? H&E

?: not specified; ♀: female; ♂: male; b.w.: body weight; ELISA: enzyme-linked immunosorbent assay; H&E: hematoxylin and eosin histological staining; qRT-PCR: real-time quantitative reverse transcription polymerase chain reaction; TUNEL: terminal deoxynucleotidyl transferase dUTP nick end labeling; AC-H3: acetilated histone H3; AC-H4: acetilated histone H4; AKT: protein kinase B; P-AKTSer473: phosphorylated AKT serine 473; Bcl-2: B-cell lymphoma 2; BAX: Bcl-2 associated X protein; CD4: cluster of cuadruple differentiation; CD8: cluster of differentiation 8; CD19: cluster of differentiation 19; CD31: cluster of differentiation 31; CD34: hematopoietic progenitor cell antigen CD34; CDK4: cyclin-dependent kinase 4; P21: cyclin-dependent kinase inhibitor 1; P27: cyclin-dependent kinase inhibitor 1B (CDKN1B); P-ERK: phosphorylated extracellular signal-regulated kinases; Plk1: polo-like kinase 1; Chk1: checkpoint kinase 1; cTnT: cardiac troponin T; CYP: cytochrome P450; DOX: doxorubicin; DP1: transcriptional factor DP1; E2F1: transcriptional factor E2F1; Hes1: transcription factor HES1; FGF-2: fibroblast growth factor; VEGF: vascular endothelial growth factor; IL-2: interleukin 2; Ki-67: Ki-67 antigen; MDR1: multidrug resistance protein 1; MMP9: matrix metallopeptidase 9; NF-κB: nuclear factor kappa-light-chain-enhancer of activated B cells; Notch 1: Notch homolog 1; P53: P53 protein; P65: nuclear factor NF-kappa-B p65 subunit; P-Rb: retinoblastoma protein; PTEN: phosphatase and tensin homolog; RBP-jk: recombining binding protein suppressor of hairless; STAT3: signal transducer and activator of transcription 3; TNF-α: tumor necrosis factor-alpha; Tyr 705-P-STAT3: phosphorylated signal transducer and activator of transcription 3; CAT: catalase; LPO: lipid peroxidation; MDA: malondialdehyde; SOD: superoxide anion; SOD1: superoxide dismutase 1; T-AOC: total antioxidant capacity; TBARS: thiobarbituric acid reactive substances; AFP: alpha-fetoprotein; AI: atherogenic index; ALP: alkaline phosphatase; ALT: alanine aminotransferase; AST: aspartate aminotransferase; BUN: blood urea nitrogen; CK-MB: creatinine kinase-MB; CK: creatinine kinase; G6PD: glucose-6-phosphate dehydrogenase; GPx: glutathione peroxidase; GPT: glutamic-pyruvic transaminase; γ-GT: gamma-glutamyl transferase; GR: glutathione reductase; GSH: reduced glutathione; GST: glutathione transferase; HDL-Ch: high-density lipoprotein cholesterol; LDL: low-density lipoprotein cholesterol; VLDL: very low-density lipoprotein cholesterol; HSPGs: heparan sulfate proteoglycans; LASA: lipid associated sialic acid; LDH: lactate dehydrogenase; MPO: hepatic myeloperoxidase; PrPr-SHs: protein thiols; TBARS: thiobarbituric acid reactive substances; T-Ch: triglyceride; TSA: total sialic acid; T-SH: total thiol; NPSH: nonprotein thiol.