Search

Background & Aims: Squamous cell carcinoma antigen recognized by T cells 3 (SART3), a tumor-associated antigen expressed in many cancers, functions in tumor rejection. In this study, we investigated its usefulness as an immunotherapeutic target in hepatocellular carcinoma (HCC). Methods: The expression of SART3 in hepatoma cell lines and HCC tissues was investigated by immunofluorescence and immunohistochemical analyses. Two peptides derived from SART3 (SART3109 and SART3315) were used for immunological analysis. T-cell responses were investigated by interferon-gamma (IFN-γ) enzyme-linked immunospot and cytotoxic T lymphocyte (CTL) assays using peripheral blood mononuclear cells (PBMCs) in 47 patients, and tumor-infiltrating lymphocytes in 8 of 47 patients with HCC. The safety of immunotherapy using a SART3-derived peptide was investigated by vaccinations of SART3109 in 12 patients with HCC (trial registration: UMIN000005677). Results: The immunofluorescence and immunohistochemical analyses showed that SART3 was expressed in six HCC cell lines, and in HCC tissues including of alpha-fetoprotein-negative individuals. SART3-specific CTLs were generated by stimulating PBMCs with the peptides, and they showed cytotoxicity against HCC cells expressing the protein. Of the 47 HCC patients, 25.5% and 10.6% showed significant responses to SART3109 and SART3315, respectively. The infiltration of SART3109-specific IFN-γ-producing CTLs into the tumor site was confirmed. In the vaccination study, no severe adverse events were observed, and the peptide-specific CTLs were newly induced in four of five patients tested. Conclusions: SART3 is an immunotherapeutic candidate, and peptides from this antigen may be applied in HCC immunotherapy. Trial Registration: [ABSTRACT FROM AUTHOR]

Hepatocytic stem cells (HSCs) have inhibitory effects on hepatocarcinoma cells. The present study investigated the effects of HSC activity in hepatocarcinoma cells in vitro. A Transwell co-culture system of hepatocytic precursor (stem-like) WB-F344 cells and hepatoma CBRH-7919 cells was used to assess HSC activity in metastasized hepatoma cells in vitro. Nude mouse xenografts were used to assess HSC activity in vivo. Co-culture of hepatoma CBRH-7919 cells with WB-F344 cells suppressed the growth and colony formation, tumor cell migration and invasion capacity of CBRH-7919 cells. The nude mouse xenograft assay demonstrated that the xenograft size of CBRH-7919 cells following co-culture with WB-F344 cells was significantly smaller compared with that of control cells. Furthermore, the expression levels of the epithelial markers E-cadherin and β-catenin were downregulated, while the mesenchymal markers α-SMA and vimentin were upregulated. Co-culture of CBRH-7919 cells with WB-F344 cells downregulated NF-κB and phospho-Akt expression. In conclusion, hepatocytic precursor (stem-like) WB-F344 cells inhibited the growth, colony formation and invasion capacity of metastasized hepatoma CBRH-7919 cells in vitro and in vivo by downregulating Akt/NF-κB signaling. [ABSTRACT FROM AUTHOR]

Background/Aims: Nonalcoholic fatty liver disease (NAFLD) progression to fibrosis, cirrhosis and hepatocellular carcinoma, alters the cellular composition of this organ. During late-stage NAFLD, fibrotic and possibly cancerous cells can proliferate and, like normal hepatocytes, are exposed to high concentrations of fatty acids from both surrounding tissue and circulating lipid sources. We hypothesized that primary human activated hepatic stellate cells and epithelial hepatoma (HepG2) cells respond differently to lipotoxic conditions, and investigated the mechanisms involved. Methods: Primary activated hepatic stellate cells and HepG2 cells were exposed to pathophysiological concentrations of fatty acids and comparative studies of lipid metabolic and stress response pathways were performed. Results: Both cell types remained proliferative during exposure to a combination of palmitate plus oleate reflective of the general saturated versus unsaturated fatty acid composition of western diets. However, exposure to either high palmitate or high oleate alone induced cytotoxicity in activated stellate cells, while only palmitate caused cytotoxicity in HepG2 cells. mRNA microarray and biochemical comparisons revealed that stellate cells stored markedly less fatty acids as neutral lipids, and had reduced capacity for beta-oxidation. Similar to previous observations in HepG2 cells, palmitate, but not oleate, induced ER stress and actin stress fiber formation in activated stellate cells. In contrast, oleate, but not palmitate, induced the inflammatory signal TXNIP, decreased cytoskeleton proteins, and decreased cell polarity preceding cell death in activated stellate cells. Conclusions: Palmitate-induced lipotoxicity was associated with ER stress pathways in both primary activated hepatic stellate cells and epithelial hepatoma cells, whereas high oleate caused lipotoxicity only in activated stellate cells, possibly through a distinct mechanism involving disruption of cytoskeleton components. This may have implications for optimal dietary fatty acid compositions during various stages of NAFLD. [ABSTRACT FROM AUTHOR]

Phytochemical investigation of Abies faxoniana Rehd. led to the isolation of two pairs of new epimeric spirolactone-type triterpenoids ( 1/1′ and 2/2′ ) and 11 known terpenoids ( 3 − 13 ). Compounds 1/1′ and 2/2′ were isolated as epimeric mixtures due to the C-23 ketal tautomerism in their spirolactone structures. The dynamic HPLC manifested that the C-23 epimeric mixtures interconverted into each other in solution. Structure determinations were based on extensive NMR and HRESIMS spectroscopic analysis. Meanwhile, their cytotoxic activities were tested by MTT method. Compound 5 showed cytotoxicities against MCF-7 and A549 cells with IC 50 values of 6.5 and 5.7 μM, respectively. Compounds 1/1′ had IC 50 values of 10.0 and 12.3 μM for Huh7 and SMMC7721 cells, respectively. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is extremely resistant towards pharmacological therapy. To date, the multi-kinase inhibitor Sorafenib is the only available therapeutic agent with the potential to prolong patient survival. Using the human hepatoma cell lines HepG2 and Huh7, we analyzed anti-cancer activities of 6 purified havanensin type limonoids isolated from the traditional African medicinal plant Trichilia rubescens Oliv. Our results show that two of the compounds, TR4 (TS3) and TR9 (Rubescin E) reduced hepatoma cell viability, but not primary hepatocyte viability, at TC50s of 5 to 10 μM. These were significantly lower than the TC50s for Sorafenib, the histone deacetylase inhibitor SAHA or 5-Fluoruracil. In comparison, TR3 (Rubescin D), a limonoid isolated in parallel and structurally highly similar to TR4 and TR9, did not interfere with hepatoma cell viability. Both, TR4 and TR9, but not TR3, induced apoptosis in hepatoma cells and interfered with NF-κB activation. TR4 as well as TR9 significantly supported anti-cancer activities of Sorafenib. In summary, the limonoids TR4 and TR9 exhibit anti-cancer activities and support Sorafenib effects in vitro, having the potential to support future HCC therapy. [ABSTRACT FROM AUTHOR]

During hepatitis B virus (HBV) infection, three viral envelope proteins of HBV are overexpressed in the endoplasmic reticulum (ER). The large S protein (LHBs) and truncated middle S protein (MHBst) have been documented to play roles in regulating host gene expression and contribute to hepatic disease development. As a predominant protein at the ultrastructural level in biopsy samples taken from viremic patients, the role of the middle S protein (MHBs) remains to be understood despite its high immunogenicity. When we transfected hepatocytes with an enhanced green fluorescent protein (EGFP)-tagged MHBs expressing plasmid, the results showed that expression of MHBs cause an upregulation of IL-6 at the message RNA and protein levels through activating the p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-κB) pathways. The use of specific inhibitors of the signaling pathways can diminish this upregulation. The use of BAPTA-AM attenuated the stimulation caused by MHBs. We further found that MHBs accumulated in the endoplasmic reticulum and increased the amount of glucose regulated protein 78 (GRP78/BiP). Our results provide a possibility that MHBs could be involved in liver disease progression. [ABSTRACT FROM AUTHOR]

Aims MicroRNA-505 (miR-505) expressions have been reported to be altered in the serum of HCC patients. However, the effect and underlying mechanism of miR-505 in hepatoma cells remains poorly understood. The present study intended to investigate the expression levels and the probable role and molecular basis of miR-505 in hepatoma cells. Main methods Real-time PCR was used to determine the miR-505 expressions in hepatoma cell lines QGY-7703, SMMC-7721 and MHCC97. Furthermore, an up-or down-regulation of miR-505 was performed in MHCC97 by transfected with miR-505 mimics or anti-miR-505, respectively. Cell proliferation, cell invasion, and epithelial-mesenchymal transition were determined. Moreover, the target gene of miR-505 was also investigated. Key findings The expressions of miR-505 were down-regulated in three hepatoma cell lines. MHCC97 possessed the lowest miR-505 levels among the three hepatoma cell lines. Furthermore, the up-regulation of miR-505 suppressed, whereas the down-regulation of miR-505 promoted proliferation, invasion and epithelial-mesenchymal transition in MHCC97. Moreover, miR-505 could directly bind to the 3′-untranslated region of High-Mobility Group Box 1. Notably, High-Mobility Group Box 1 knockdown apparently promoted cell proliferation and invasion in MHCC97. Significance We investigated that MiR-505 regulates proliferation and invasion in MHCC97 cells via targeting High-Mobility Group Box 1. [ABSTRACT FROM AUTHOR]

Hepatitis C virus (HCV) induces interferon (IFN) stimulated genes in the liver despite of distinct innate immune evasion mechanisms, suggesting that beyond HCV infected cells other cell types contribute to innate immune activation. Upon coculture with HCV replicating cells, human CD141+ myeloid dendritic cells (DC) produce type III IFN, whereas plasmacytoid dendritic cells (pDC) mount type I IFN responses. Due to limitations in the genetic manipulation of primary human DCs, we explored HCV mediated stimulation of murine DC subsets. Coculture of HCV RNA transfected human or murine hepatoma cells with murine bone marrow-derived DC cultures revealed that only Flt3-L DC cultures, but not GM-CSF DC cultures responded with IFN production. Cells transfected with full length or subgenomic viral RNA stimulated IFN release indicating that infectious virus particle formation is not essential in this process. Use of differentiated DC from mice with genetic lesions in innate immune signalling showed that IFN secretion by HCV-stimulated murine DC was independent of MyD88 and CARDIF, but dependent on TRIF and IFNAR signalling. Separating Flt3-L DC cultures into pDC and conventional CD11b-like and CD8α-like DC revealed that the CD8α-like DC, homologous to the human CD141+ DC, release interferon upon stimulation by HCV replicating cells. In contrast, the other cell types and in particular the pDC did not. Injection of human HCV subgenomic replicon cells into IFN-β reporter mice confirmed the interferon induction upon HCV replication in vivo. These results indicate that HCV-replicating cells stimulate IFN secretion from murine CD8α-like DC independent of infectious virus production. Thus, this work defines basic principles of viral recognition by murine DC populations. Moreover, this model should be useful to explore the interaction between dendritic cells during HCV replication and to define how viral signatures are delivered to and recognized by immune cells to trigger IFN release. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma is associated with high mortality, and tumor metastasis is an important reason for poor prognosis. However, metastasis has not been effectively prevented in clinical therapy and the mechanisms underlying metastasis have not been fully characterized. CCAAT/enhancer-binding protein-α (C/EBPα) is a transcriptional regulator with an essential role in tumor metastasis. We used short-activating RNAs (saRNA) to enhance expression of C/EBPα. Intravenous injection of C/EBPα-saRNA in a nude mouse liver orthotopic xenograft tumor model inhibited intrahepatic and distant metastasis. C/EBPα-saRNA-treated mice showed increased serum levels of albumin and decreased alanine aminotransferase (ALT), glutamic-oxalacetic transaminase (AST), indicating a role of C/EBPα in improving liver function. Migration and invasion were inhibited in hepatoma cell lines transfected with C/EBPα-saRNA. We also observed an inhibition of epithelial-mesenchymal transition (EMT) and suppression of epidermal growth factor receptor (EGFR), EGFR phosphorylation, and β-catenin in C/EBPa-saRNA-transfected cells. Our results suggested that C/EBPα-saRNA successfully inhibited HCC metastasis by inhibiting EGFR/β-catenin signaling pathway mediated EMT in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Based on the scaffold hybridization strategy, twenty-four indole-guanidines were designed and synthesized. Subsequently, anti-proliferative activity against various cancer cells indicated that most of these hybrids exhibited moderate to high anti-proliferative activity, especially for human hepatoma cell lines. Selectivity investigation showed that these hybrids showed the best selectivity for SMMC-7721 subtype in human hepatoma cells. Particularly, (E)-3-((2-(N -pentylcarbamimidoyl)hydrazono)methyl)-1 H -indol-5-yl 4-methylbenzoate (19) and (E)-3-((2-(N -pentylcarbamimidoyl)hydrazono)methyl)-1 H -indol-5-yl 4-methoxybenzoate (22) exhibited potent inhibition against SMMC-7721 cells with IC 50 values of 0.057 μM and 0.042 μM, respectively, far outperforming that of Sorafenib. Meanwhile, hybrids 19 and 22 exhibited no significant cytotoxicity against normal cells such as HEK293 cells and HEK293T cells. Moreover, further investigations indicated that hybrid 22 effectively induced apoptosis, arrested the cell cycle at S phase, and selectively down regulated expression of p-STAT3, JAK2 and BRAF in SMMC-7721 cells in a dose-dependent manner. Molecular docking indicated that hybrid 22 exhibited high affinity with STAT3 and BRAF. In summary, hybrid 22 was developed as a potential and effective anti-hepatoma candidate, which was worthy of further investigation. Discovery of several indole-guanidine hybrids as novel anti-cancer agents with novel scaffold. [Display omitted] • Discovery of indole-guanidine hybrids as novel and potent anti-cancer candidates with novel scaffold. • Two hybrids exhibited potent and selective anti-proliferative activities with no significant cytotoxicity. • The candidate 22 could induce apoptosis, arrest the cell cycle, down regulate p-STAT3, JAK2 and BRAF. • In silico study indicated that the candidate 22 showed high affinity with STAT3 and BRAF. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is one of the most common cancers in Taiwan. Although chemotherapy is the primary treatment for HCC patients, drug resistance often leads to clinical failure. Galectin-1 is a beta-galactoside binding lectin which is up-regulated in HCC patients and promotes tumor growth by mediating cancer cell adhesion, migration and proliferation, but its role in chemoresistance of HCC is poorly understood. In this study we found that galectin-1 is able to lead to chemoresistance against cisplatin treatment, and subsequent inhibition has reversed the effect of cell death in HCC cells. Moreover, galectin-1 was found to induce autophagic flux in HCC cells. Inhibition of autophagy by inhibitors or knockdown of Atg5 cancels galectin-1-induced cisplatin resistance in HCC cells. Increase of mitophagy triggered by galectin-1 was found to reduce the mitochondrial potential loss and apoptosis induced by cisplatin treatment. Finally, using an in situ hepatoma mouse model, we clearly demonstrated that inhibition of galectin-1 by thiodigalactoside could significantly augment the anti-HCC effect of cisplatin. Taken together, our findings offer a new insight into the chemoresistance galectin-1 causes against cisplatin treatment, and points to a potential approach to improve the efficacy of cisplatin in the treatment of HCC patients. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the effect of hepatitis B virus X protein (HBx) on the expression of DNA methyltransferase (DNMT)3A/3B and suppressors of cytokine signaling-1 (SOCS-1), as well as promoter CpG island methylation of the SOCS-1 gene. Stable hepatocyte cell lines expressing the HBx gene (pcDNA-X/QSG7701) or an empty gene (pcDNA3.0/QSG7701) were established. Reverse transcription quantitative polymerase chain reaction (PCR) was used to detect the mRNA expression levels of DNMT3A/3B and SOCS-1. Immunohistochemistry was used to detect the protein expression of DNMT3A/3B. Methylation-specific PCR (MSP) was used to detect the methylation status of the SOCS-1 gene promoter. The mRNA and protein expression levels of DNMT3A/3B were significantly higher in the pcDNA-X/QSG7701-transfected cells, compared with those in the pcDNA3.0/QSG7701 or non-transfected QSG7701 cells (P<0.05), whereas the relative mRNA expression of SOCS-1 was significantly lower in the pcDNA-X/QSG7701 cells compared with the pcDNA3.0/QSG7701 and non-transfected QSG7701 cells (F=19.6; P<0.05). Western blot analysis showed that the protein expression of SOCS-1 was significantly lower in the pcDNA-X/QSG7701 cells, compared with the pcDNA3.0/QSG7701 or non-transfected QSG7701 cells (F=19.4; P<0.05). The results of the MSP analysis showed that SOCS-1 promoter region methylation was present only in the pcDNA-X/QSG7701 cells. The HBV-X gene upregulated the mRNA and protein expression levels of DNMT3A/3B, downregulated the expression of SOCS-1 and increased SOCS-1 gene promoter CpG island methylation. This may provide a potential explanation of the mechanism underlying HBx-associated hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

Background: Hepatitis B virus (HBV)-X protein(HBx) is a transactivator of host several cellular genes including alpha-fetoprotein(AFP) and AFP receptor(AFPR) which contributes to HBV-associated tumor development. The expression of AFP/AFPR are correlated with hepatocellular carcinoma(HCC)-initial cells. But the role of AFP and AFPR in promoting occurrence of HBV-related HCC were still unclear. Methods: A total of 71 clinical patients' liver specimens, normal human liver cells L-02 and HCC cell lines, PLC/PRF/5 were selected for analyzing the effects of HBx on expression of AFP, AFPR and Src. The expression of goal proteins were detected by Immunohistochemical stained and Western blotting; HBx-expressed vectors were constructed and transfected into L-02 cells, laser confocal microscopy was applied to observe expression and location of AFP, AFPR and Src in the normal liver cells and HCC cells, soft agar colony formation assay was used to observe colonies formed of the cells. Results: We confirmed HBx gives preference to promote the expression of AFP and AFPR; HBx priors to up-regulate the expression of AFPR and AFP in L-02 cells and in normal liver specimens; AFPR signal been able to stimulate Src expression. The results also indicated that phosphatidylinositol 3-kinase(PI3K) inhibitors Ly294002 and GDC0941 effectively suppress AFPR mediated up-regulation expression of Src in AFPR positive HCC lines. Conclusions: HBx priors to drive the expression of AFP and AFPR to promote expression of Src in normal liver cells and hepatoma cells; AFP and AFPR maybe play pivotal role in HBV-related hepatocarcinogenesis; Targeting AFPR is an available therapeutic strategy of HCC. [ABSTRACT FROM AUTHOR]

Apolipoprotein[a] (apo[a]) is a highly polymorphic glycoprotein that forms a covalent complex with apolipoprotein B-100 (apoB-100), producing a lipoprotein species referred to as lipoprotein[a] (Lp[a]). We have studied the effects of alterations in glycosylation of apo[a] on its intracellular processing and secretion as well as its ability to associate with low density lipoprotein (LDL) apoB-100. HepG2 cells transfected with a 6 kringle IV (6 K-IV) apo[a] minigene were treated with tunicamycin, an inhibitor of N-linked glycosylation, which eliminated apo[a]-B-100 complexes from the media. Tunicamycin treatment also reduced secretion of the 6 K-IV apo[a] protein from transfected McA-RH7777 cells by ∼50%, but completely eliminated secretion of apo[a] species containing 9 and 17 K-IV repeats. Mixing experiments, performed with radiolabeled media (±tunicamycin) from transfected McA-RH7777 cells, demonstrated no alteration in the extent of association of apo[a] with human LDL. Similar mixing experiments using culture media from glycosylation-defective mutant chinese hamster ovary (CHO) cells transfected with the same apo[a] minigene showed identical results. Apo[a] secretion was demonstrated in all mutant cell lines in the absence of either N- or O-linked (or both) glycosylation. The mechanisms underlying the reduced secretion of apo[a] from transfected hepatoma cells were examined by pulse-chase radiolabeling and apo[a] immunoprecipitation. Tunicamycin treatment altered the efficiency of precursor apo[a] processing from the ER by increasing its ER retention time. The increased accumulation of precursor apo[a] in the ER was associated with alterations in the kinetics of association with two resident endoplasmic reticulum (ER) chaperone proteins, calnexin and BiP. These findings suggest that the glycosylation state and size of apo[a] appear to play a role in regulating its efficient exit from the endoplasmic reticulum. However, neither N- nor O-linked glycosylation of apo[a] exerts a major regulatory role in its covalent association with apoB-100.—Bonen, D. K., F. Nassir, A. M. L. Hausman, and N. O. Davidson. Inhibition of N-linked glycosylation results in retention of intracellular apo[a] in hepatoma cells, although nonglycosylated and immature forms of apolipoprotein[a] are competent to associate with apolipoprotein B-100 in vitro.

Background: The polysaccharide extract of C. sinensis, Isaria felina (IF), has antitumor effects. Selenium (Se) can improve disease prevention and reduce the toxicity of toxic elements, but the effect of Se-enriched IF on hepatoma remains unknown., Objective: To determine the organic transformation of Se and compare the antitumor effects between Se-enriched IF (IF-Se) and IF on xenograft H22 hepatoma-bearing mice., Methods: Se was added to the solid-state culture medium, and the organic Se content was detected by HPLC-ICP-MS. Forty-two Kunming mice were randomly divided into seven groups to test the antitumor effects of low- (300 mg/kg) and high- (600 mg/kg) doses of IF-Se and IF through xenograft. Huai'er granules were administered as the positive control. In addition, interleukin (IL)-2 and vascular endothelial growth factor (VEGF) expressions were measured by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry method., Results: The conversion rate in the IF-Se70, IF-Se140, and IF-Se280 groups were 91.5%, 93.4%, and 89.3%, respectively. Therefore, IF-Se140 was used to carry out the subsequent experiments. The tumor inhibition rates of IF-Se were significantly higher compared with IF ( P < 0.05). Moreover, the spleen coefficient, IL-2, and VEGF expression levels significantly decreased (all Ps < 0.05), and the thymus coefficient significantly increased ( P < 0.05) in the high-dose IF-Se group compared with the model control group., Conclusion: The inhibitory effects of IF on H22 hepatoma-bearing mice were enhanced after Se enrichment. Therefore, Se-enriched IF might be a new strategy for treating hepatoma., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Sorafenib (Nexavar) is currently the only FDA-approved small molecule targeted therapy for advanced hepatocellular carcinoma. The use of structural analogues and derivatives of sorafenib has enabled the elucidation of critical targets and mechanism(s) of cell death for human cancer lines. We previously performed a structure-activity relationship study on a series of sorafenib analogues designed to investigate the inhibition overlap between the major targets of sorafenib Raf-1 kinase and VEGFR-2, and an enzyme shown to be a potent off-target of sorafenib, soluble epoxide hydrolase. In the current work, we present the biological data on our lead sorafenib analogue, t-CUPM, demonstrating that this analogue retains cytotoxicity similar to sorafenib in various human cancer cell lines and strongly inhibits growth in the NCI-60 cell line panel. Co-treatment with the pan-caspase inhibitor, Z-VAD-FMK, failed to rescue the cell viability responses of both sorafenib and t-CUPM, and immunofluorescence microscopy shows similar mitochondrial depolarization and apoptosis-inducing factor release for both compounds. These data suggest that both compounds induce a similar mechanism of caspase-independent apoptosis in hepatoma cells. In addition, t-CUPM displays anti-proliferative effects comparable to sorafenib as seen by a halt in G0/G1 in cell cycle progression. The structural difference between sorafenib and t-CUPM significantly reduces inhibitory spectrum of kinases by this analogue, and pharmacokinetic characterization demonstrates a 20-fold better oral bioavailability of t-CUPM than sorafenib in mice. Thus, t-CUPM may have the potential to reduce the adverse events observed from the multikinase inhibitory properties and the large dosing regimens of sorafenib. [ABSTRACT FROM AUTHOR]

Chronic hepatitis B virus (HBV) and hepatitis D virus (HDV) infections are major causes of liver disease and hepatocellular carcinoma worldwide. Despite the presence of an efficient preventive vaccine, more than 250 million patients are chronically infected with HBV. Current antivirals effectively control but only rarely cure chronic infection. While the molecular biology of the two viruses has been characterized in great detail, the absence of robust cell culture models for HBV and/or HDV infection has limited the investigation of virus-host interactions. Native hepatoma cell lines do not allow viral infection, and the culture of primary hepatocytes, the natural host cell for the viruses, implies a series of constraints restricting the possibilities of analyzing virus-host interactions. Recently, the discovery of the sodium taurocholate co-transporting polypeptide (NTCP) as a key HBV/HDV cell entry factor has opened the door to a new era of investigation, as NTCP-overexpressing hepatoma cells acquire susceptibility to HBV and HDV infections. In this review, we summarize the major cell culture models for HBV and HDV infection, discuss their advantages and limitations and highlight perspectives for future developments.

The tumor suppressor gene CD82, also known as KAI1, may act as a general suppressor of metastasis in numerous types of cancer. It is hypothesized that downregulation of CD82 gene expression may be an important factor in the induction of hepatocellular carcinoma (HCC), however the mechanism for this requires further study. In the present study, the relative mRNA and protein expression levels of the CD82 gene were determined in HCC and adjacent non-tumor tissues. The association between the CD82 gene and the hepatitis B virus (HBV) was also investigated, by quantitative polymerase chain reaction, western blotting, luciferase reporter assays and mass spectrometry with matrix-assisted laser desorption/ ionization time-of-flight mass array. CD82 expression was shown to be suppressed in response to HCC promoter methylation. Relative CD82 mRNA and protein expression levels were downregulated in HCC tissues (P<0.05). HBx protein inhibited CD82 promoter activity and subsequently the mRNA and protein expression levels. Furthermore, it was demonstrated that HBV could inhibit the expression of CD82 at the transcriptional level, and repress the activity of the CD82 promoter through hypermethylation. In addition, the methyl enzyme inhibitor 5-aza-CdR could induce the CD82 promoter activity and the relative expression level of CD82 mRNA, as observed by an increase in luciferase activity driven by the CD82 promoter. The observations of the present study suggest that hypermethylation of the CD82 promoter may be an event leading to the development of HCC. Low expression of CD82 is likely to be involved in tumor progression. HBV may inhibit the expression of CD82 through hypermethylation of the promoter in hepatoma cells. [ABSTRACT FROM AUTHOR]

Celecoxib has been shown to have antitumor effect in previous studies but the mechanisms are unclear. The effect of celecoxib on cytosolic Ca2+ concentrations ([Ca2+]i) and viability in HA59T human hepatoma cells was explored. The Ca2+-sensitive fluorescent dye fura-2 was applied to measure [Ca2+]i. Celecoxib at concentrations of 10–50 μM induced a [Ca2+]i rise in a concentration-dependent manner. The response was reduced by 80% by removing Ca2+. Celecoxib induced Mn2+ influx, leading to quenching of fura-2 fluorescence. Celecoxib-evoked Ca2+ entry was suppressed by nifedipine, econazole, SK&F96365, and protein kinase C modulators. In the absence of extracellular Ca2+, incubation with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin nearly abolished celecoxib-induced [Ca2+]i rise. Incubation with celecoxib abolished thapsigargin-induced [Ca2+]i rise. Inhibition of phospholipase C with U73122 abolished celecoxib-induced [Ca2+]i rise. At 1–50 μM, celecoxib inhibited cell viability by less than 20%, which was not reversed by chelating cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N, N, N′, N′-tetraacetic acid/acetoxy methyl (BAPTA/AM). Celecoxib (10–50 μM) also induced apoptosis. In sum, in HA59T hepatoma cells, celecoxib induced a [Ca2+]i rise by evoking phospholipase C-dependent Ca2+ release from the endoplasmic reticulum and Ca2+ entry via protein kinase C-sensitive store-operated Ca2+ channels. Celecoxib also caused cell death via apoptosis. [ABSTRACT FROM PUBLISHER]

Copper ions are known to induce insulin-like effects in various cell lines, stimulating the phosphoinositide 3'-kinase (PI3K)/Akt signaling cascade and leading to the phosphorylation of downstream targets, including FoxO transcription factors. The aim of this work was to study the role of insulin- and IGF1-receptors (IR and IGF1R) in insulin-like signaling induced by copper in HepG2 human hepatoma cells. Cells were exposed to Cu(II) at various concentrations for up to 60 min. While Akt and FoxO1a/FoxO3a were strongly phosphorylated in copper- and insulin-treated cells at all time points studied, only faint tyrosine phosphorylation of IR/IGF1R was detected in cells exposed to Cu(II) by either immunoprecipitation/immunoblot or by immunoblotting using phospho-specific antibodies, whereas insulin triggered strong phosphorylation at these sites. Pharmacological inhibition of IR/IGF1R modestly attenuated Cu-induced Akt and FoxO phosphorylation, whereas no attenuation of Cu-induced Akt activation was achieved by siRNA-mediated IR depletion. Cu(II)-induced FoxO1a nuclear exclusion was only slightly impaired by pharmacological inhibition of IR/IGF1R, whereas insulin-induced effects were blunted. In contrast, genistein, a broad-spectrum tyrosine kinase inhibitor, at concentrations not affecting IR/IGF1R, attenuated Cu(II)-induced Akt phosphorylation, pointing to the requirement of tyrosine kinases other than IR/IGF1R for Cu(II)-induced signaling. [ABSTRACT FROM AUTHOR]

To gain a better understanding of the anticancer effects of hydroxyapatite (HAP) nanoparticles in vivo and in vitro, the effects of the interaction of HAP nanoparticles with hepatoma cells were explored. HAP nanoparticles were prepared by homogeneous precipitation and characterized by laser particle analysis and transmission electron microscopy (TEM). HAP nanoparticles were observed to be uniformly distributed, with rod-like shapes and diameters in the range of 42.1-87.1 nm. Overnight attached, suspended, and proliferating Bel-7402 cells were incubated with HAP nanoparticles. Inverted microscopy observation revealed that HAP nanoparticles with a cell membrane showed good adsorption. TEM demonstrated that HAP nanoparticles were present on the surface of cells, continuously taken up by cells through endocytosis, and transported in vesicles close to the nucleus. Fluorescence microscopy showed that the concentrations of intracellular Ca labeled with Fluo-3 calcium fluorescent probe were significantly enhanced. In addition, inverted microscopy observation revealed that suspended cells treated with HAP nanoparticles did not adhere to the culture bottle, resulting in cell death. After the overnight attached cells were treated with HAP nanoparticles for 96 h with increasing doses of HAP nanoparticles, inverted microscopy observation revealed that cell proliferation was slowed and cell-cell adhesion was weakened. Feulgen staining and image analysis indicated that the nuclear DNA content of the cells was markedly reduced, and argyrophilic nucleolar organizer region (AgNOR) staining and image analysis indicated that the number of AgNORs was significantly decreased. Therefore, hepatoma cells brought about the adsorption, uptake, transport and degradation of HAP nanoparticles. In addition, HAP nanoparticles affected hepatoma cells with regard to cell-cell adhesion, cell and extracellular matrix adhesion, and DNA and protein synthesis; thus inhibiting cell proliferation. This understanding of the effects of interaction between HAP nanoparticles and hepatoma cells is useful for further study of the anticancer mechanisms of HAP nanoparticles. [ABSTRACT FROM AUTHOR]

Highlights: [•] Inhibition of H2S/CSE pathway strongly stimulates cellular apoptosis. [•] Inhibition of H2S/CSE pathway suppresses cell growth by blocking EGFR pathway. [•] H2S/CSE pathway is critical for maintaining the proliferation of hepatoma cells. [ABSTRACT FROM AUTHOR]

ABSTRACT Arsenic (As) is considered a top environmental chemical of human health because it has been linked to adverse health effects including cancer, diabetes, cardiovascular disease, and reproductive and developmental problems. In several cell culture and animal models, As acts as an endocrine disruptor, which may underlie many of its health effects. Previous work showed that steroid receptor (SR)-driven gene expression is disrupted in cells treated with inorganic As (arsenite, iAs+3). In those studies, low iAs+3 concentrations (0.1-0.7 μM) stimulated hormone-inducible transcription, whereas somewhat higher but still non-cytotoxic levels (1-3 μM) inhibited transcription. This investigation focuses on the mechanisms underlying these inhibitory effects and evaluates the role of methylated trivalent As metabolites on SR function. Recent evidence suggests that, compared with iAs, methylated forms may have distinct biochemical effects. Here, fluorescence polarization (FP) experiments utilizing purified, hormone-bound human glucocorticoid (GR) and progesterone receptor (PR) have demonstrated that neither inorganic (iAs+3) nor dimethylated (DMA+3) species of trivalent As affect receptor interactions with glucocorticoid DNA response elements (GREs). However, monomethylated forms (monomethylarsenite, MMA+3 and monomethylarsonic diglutathione, MADG) strongly inhibit GR-GRE and PR-GRE binding. Additionally, speciation studies of iAs+3-treated H4IIE rat hepatoma cells show that, under treatment conditions that cause inhibition of hormone-inducible gene transcription, the intracellular concentration of MADG is sufficient to inhibit GR-GRE and PR-GRE interactions in vivo. These results indicate that arsenic's inhibitory endocrine disruption effects are probably caused in part by methylated metabolites' disruption of SR ability to bind DNA response elements that are crucial to hormone-driven gene transcription. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Purpose: The radiation-induced bystander effect (RIBE) has important implications for the efficiency of radiotherapy but the underlying role of cellular metabolism is widely unknown. The roles of synthesis of cytochrome c oxidase 2 (SCO2), a key effector for respiratory chain, and related signaling factors in α-particle-induced bystander damage were currently investigated in a liver cell co-culture system. Materials and methods: Human hepatoma cells of HepG2 with wild-type p53 (wtp53) and Hep3B (p53 null) were irradiated with 0.4 Gy of α-particles and co-cultured with non-irradiated normal liver cells HL-7702 for 6 h, then the incidence of micronucleus (MN) in the bystander HL-7702 cells was analyzed. The expressions of total P53, phospho-P53 (p-P53), SCO2, and reactive oxygen species (ROS) in the irradiated hepatoma cells were detected. In some experiments, the hepatoma cells were respectively treated with p53 siRNA, SCO2 siRNA, or dimethyl sulfoxide (DMSO) before irradiation. Results: Bystander damage in HL-7702 cells was induced by α-irradiated HepG2 cells but not by α-irradiated Hep3B cells, and this bystander effect was diminished when the irradiated HepG2 cells were pretreated with p53 siRNA, SCO2 siRNA, or DMSO. Meanwhile, the expressions of p-P53 protein and SCO2 mRNA, the activity of SCO2 protein, and intracellular ROS were all increased in the irradiated HepG2 cells but not Hep3B cells and these expressions were eliminated by p53 siRNA treatment. Moreover, the radiation-enhanced expressions of SCO2 and ROS were inhibited by SCO2 siRNA. Conclusion: α-particle-induced bystander effect was regulated by p53 and its downstream SCO2 in the irradiated hepatoma cells, and ROS generation could be an early event for triggering this bystander response. [ABSTRACT FROM AUTHOR]

[Objective] To preliminarily quantify the control efficacy of human liver cancer by propolis through evaluating the inhibitory effect on HCC cell proliferation and fat deposition. [Methods] The proliferation capacity of propolis-treated hepatoma cells was evaluated by MIT. Oleic acid induction method was adopted to delect the degree of fat deposition in HCC cell. [Results] The content of flavonoids in propolis was determined by HLPC-MS to be (191.472 ± 1.986) mg/g. The tg 9 group (propolis: olive oil = 1:2, V/V) revealed the strongest inhibitory effect on cell proliferation. The triglyceride (TG) contents in all treatment groups were in the range of 410.333 -459.672 nmol/L. [Conclusions] Therefore, propolis can not control fat deposition in cells and the preventative effect of propolis on fatty liver is not satisfactory. However, it has strong inhibitory effect on the proliferation of liver cancer cells. [ABSTRACT FROM AUTHOR]

Abstract: Signaling events triggered by interferon alpha (IFN-α) and ribavirin are involved in anti-hepatitis C virus (HCV) action. The p38 mitogen-activated protein kinase (MAPK) pathway plays an important role in HCV pathogenesis. Effects of IFN-α and ribavirin on p38 MAPK signaling were investigated in human hepatoma cells. Type I IFN receptor 2 (IFNAR2) mediated IFN-α-induced p38 MAPK phosphorylation. Also, p38 MAPK phosphorylation was enhanced by ribavirin. Treatment for 48h with a combination of IFN-α and ribavirin increased p38 MAPK phosphorylation, whereas the treatment for 72h reduced p38 MAPK phosphorylation. Cell culture-derived HCV (HCVcc) infection dramatically increased p38 MAPK phosphorylation and such phosphorylation was inhibited by IFN-α or ribavirin. Moreover, siRNA-mediated knockdown of p38 MAPK resulted in enhancement of ribavirin-dependent HCV RNA replication. These results suggest that regulation of p38 MAPK signaling by IFN-α and ribavirin might contribute to anti-HCV action. [Copyright &y& Elsevier]

Abstract: As differentiated cells, hepatocytes primarily metabolize glucose for ATP production through oxidative phosphorylation of glycolytic pyruvate, whereas proliferative hepatocellular carcinoma (HCC) cells undergo a metabolic shift to aerobic glycolysis despite oxygen availability. Keratins, the intermediate filament (IF) proteins of epithelial cells, are expressed as pairs in a lineage/differentiation manner. Hepatocyte and HCC (hepatoma) cell IFs are made solely of keratins 8/18 (K8/K18), thus providing models of choice to address K8/K18 IF functions in normal and cancerous epithelial cells. Here, we demonstrate distinctive increases in glucose uptake, glucose-6-phosphate formation, lactate release, and glycogen formation in K8/K18 IF-lacking hepatocytes and/or hepatoma cells versus their respective IF-containing counterparts. We also show that the K8/K18-dependent glucose uptake/G6P formation is linked to alterations in hexokinase I/II/IV content and localization at mitochondria, with little effect on GLUT1 status. In addition, we find that the insulin-stimulated glycogen formation in normal hepatocytes involves the main PI-3 kinase-dependent signaling pathway and that the K8/K18 IF loss makes them more efficient glycogen producers. In comparison, the higher insulin-dependent glycogen formation in K8/K18 IF-lacking hepatoma cells is associated with a signaling occurring through a mTOR-dependent pathway, along with an augmentation in cell proliferative activity. Together, the results uncover a key K8/K18 regulation of glucose metabolism in normal and cancerous hepatic cells through differential modulations of mitochondrial HK status and insulin-mediated signaling. [Copyright &y& Elsevier]

Primary liver cancer is one of the highest mortality malignant tumors in the world. China is a high occurrence area of primary liver cancer. Diagnosis of liver cancer, especially early diagnosis, is essential for improving patients' survival. Light scattering and measuring method is an emerging technology developed in recent decades, which has attracted a large number of biomedical researchers due to its advantages, such as fast, simple, high accuracy, good repeatability, and non-destructive. The hypothesis of this project is that there may be some different light scattering information between hepatoma cells and hepatocyte. Combined with the advantages of the dynamic light scattering method and the biological cytology, an experimental scheme to measure the light scattering information of cells was formulated. Hepatoma cells and hepatic cells were irradiated by a semiconductor laser (532 nm). And the Brookhaven BI-200SM wide-angle light scattering device and temperature control apparatus were adopted. The light scattering information of hepatoma cells and hepatic cells in vitro within the 15°C to 30°C temperature range was processed by a BI-9000AT digital autocorrelator. The following points were found: (a) the scattering intensities of human hepatic cells and hepatoma cells are nearly not affected by the temperature factor, and the former is always greater than the latter and (b) the relaxation time of hepatoma cells is longer than that of hepatic cells, and both the relaxation time are shortened with increasing temperature from 15°C to 25°C. It can be concluded that hepatoma cells could absorb more incident light than hepatic cells. The reason may be that there exists more protein and nucleic acid in cancerous cells than normal cells. Furthermore, based on the length relaxation time, a conclusion can be inferred that the Brownian movement of cancer cells is greater. [ABSTRACT FROM AUTHOR]

Cyclophilin, a cytosolic receptor for the immunosuppressive drug cyclosporin A, plays a role in diverse pathophysiologies along with its receptor, CD147. Although the interaction between cyclophilin A and CD147 is well established in inflammatory disease, that of cyclophilin B (CypB) with CD147 has not been fully explored, especially in cancer cell biology, and the exact molecular mechanism underlying such an association is poorly understood. In this study, we first identified high expression levels of CypB in 54 % of hepatocellular carcinoma patient tissues but in only 12.5 % of normal liver tissues. Then, we demonstrated that CypB overexpression protects human hepatoma cells against oxidative stress through its binding to CD147; this protective effect depends on the peptidyl prolyl isomerase activity of CypB. siRNA-mediated knockdown of CypB expression rendered hepatoma cells more vulnerable to ROS-mediated apoptosis. Furthermore, we also determined that a direct interaction between secreted CypB and CD147 regulates the extracellular signal-regulated kinase intracellular signaling pathway and is indispensible for the protective functions of CypB. For the first time, we demonstrated that CypB has an essential function in protecting hepatoma cells against oxidative stress through binding to CD147 and regulating the ERK pathway. [ABSTRACT FROM AUTHOR]

Abstract: Although there is in vivo evidence suggesting a role for glutathione in the metabolism and tissue distribution of vitamin C, no connection with the vitamin C transport systems has been reported. We show here that disruption of glutathione metabolism with buthionine-(S,R)-sulfoximine (BSO) produced a sustained blockade of ascorbic acid transport in rat hepatocytes and rat hepatoma cells. Rat hepatocytes expressed the Na+-coupled ascorbic acid transporter-1 (SVCT1), while hepatoma cells expressed the transporters SVCT1 and SVCT2. BSO-treated rat hepatoma cells showed a two order of magnitude decrease in SVCT1 and SVCT2 mRNA levels, undetectable SVCT1 and SVCT2 protein expression, and lacked the capacity to transport ascorbic acid, effects that were fully reversible on glutathione repletion. Interestingly, although SVCT1 mRNA levels remained unchanged in rat hepatocytes made glutathione deficient by in vivo BSO treatment, SVCT1 protein was absent from the plasma membrane and the cells lacked the capacity to transport ascorbic acid. The specificity of the BSO treatment was indicated by the finding that transport of oxidized vitamin C (dehydroascorbic acid) and glucose transporter expression were unaffected by BSO treatment. Moreover, glutathione depletion failed to affect ascorbic acid transport, and SVCT1 and SVCT2 expression in human hepatoma cells. Therefore, our data indicate an essential role for glutathione in controlling vitamin C metabolism in rat hepatocytes and rat hepatoma cells, two cell types capable of synthesizing ascorbic acid, by regulating the expression and subcellular localization of the transporters involved in the acquisition of ascorbic acid from extracellular sources, an effect not observed in human cells incapable of synthesizing ascorbic acid. [Copyright &y& Elsevier]

Hepatocellular carcinoma (HCC) is a common malignancy and a leading cause of cancer death worldwide. Hepatitis B x-interacting protein (HBXIP), a cofactor of survivin, was originally identified by binding with the C-terminus of the HBx and negatively regulated the activity of HBx. In this study, the effect of HBXIP on the hepatoma cells-induced angiogenesis was investigated. Proliferation and migration of human umbilical vein endothelial cells (HUVECs) were detected by MTT and transwell assay, respectively. Tube formation and chick chorioallantoic membrane model were used to observe the angiogenesis. Vascular endothelial growth factor activity was assayed using ELISA kits. Western blotting was performed to examine the protein expression. Our results indicated that overexpression of HBXIP increased HepG2 cell-induced endothelial cells migration, proliferation, and angiogenesis, which may be related to increasing phosphorylation of endothelial NO synthase in HUVECs. These results suggest that HBXIP may play an important role in tumorigenesis by enhancing angiogenesis in HCC . [ABSTRACT FROM AUTHOR]

Abstract: Objectives: We investigated the effects of xeroderma pigmentosum D (XPD) on the growth of hepatoma cells and the expressions of P21, Bax, Bcl-2 and Hepatitis B virus X protein (HBx). In addition, we examined whether XPD affected the aforementioned genes via the P53 pathway. Methods: Human hepatoma cells (HepG2.2.15) were transfected with XPD expression vector, followed by incubation with Pifithrin-α (P53 inhibitor). By using RT-PCR and Western blotting, the expression levels of XPD, P53, phospho-P53 (ser-15), P21, Bax, Bcl-2 and HBx were detected. The cell cycle and the apoptosis rate were examined with flow cytometry, and the cell viability was detected by MTT. Results: Over-expression of XPD up-regulated the expressions of P53, phospho-P53 (ser-15), P21 and Bax but down-regulated the expressions of Bcl-2 and HBx. XPD inhibited the viability of HepG2.2.15 and exacerbated the apoptosis. However, the inhibition of P53 by Pifithrin-α abolished the above-mentioned effects of XPD. Conclusion: XPD could suppress growth of hepatoma cells, up-regulate the expressions of P21 and Bax, and down-regulate the expressions of Bcl-2 and HBx through the P53 pathway. There may be mutual influences among XPD, P53 and HBx that co-regulate hepatocarcinogenesis. [Copyright &y& Elsevier]

Abstract: 1-Nitropyrene (1-NP) is a nitro-polycyclic aromatic hydrocarbon (nitro-PAH) present in diesel exhaust and bound to particular matter in urban air. We show that 1-NP and the referent PAH benzo(a)pyrene (BP) induce apoptosis and a lipid accumulation dependent on cytochrome P450 1A1-metabolites in mouse hepatoma cells, whereas 1-amino-pyrene had no effect. The caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethyl ketone (Z-VAD-fmk), inhibits 1-NP-induced apoptosis, but failed to alter 1-NP-triggered lipid accumulation determined by Nile red staining. We further show that cholesterol and fatty acid contents are modified after nitro-PAH exposure and that 1-NP-induced cholesterol level is partially involved in related apoptosis. In parallel, the activity of the stearoyl-CoA desaturase 1 (SCD1), determined by fatty acid analysis, and its expression are reduced by 1-NP. The role of SCD1 in 1-NP-induced apoptosis is demonstrated in cells down-expressing SCD1, in which an increased apoptosis is observed, whereas the SCD1 overexpression elicits the opposite effects. In contrast, changes in SCD1 gene expression have no effect on the induced lipid accumulation. Moreover, 1-NP increases the activity of the AMP-dependent protein kinase (AMPK) leading to a caspase-independent apoptosis. Overall, our study demonstrates that the 1-NP-induced apoptosis is caspase- and AMPK-dependent, and is associated to a decrease of SCD1 expression which results in an alteration of lipid homeostasis. [Copyright &y& Elsevier]

Background and Objectives: Significant progress has been made in treatment of hemophilia. Exvivo gene therapy is going popular due to the capability of this method in using isogenic cells for genetic manipulation and reintroducing them into same host after proliferation. Most gene therapy techniques use viral vectors, which usually harbor a strong and non-specific promoter (e.g. CMV early promoter) for driving the downstream gene. This may be a disadvantage due to uncontrollable nature of gene expression. In addition, considering the potentials of recently introduced stem cells as reservoirs and their potential to differentiate to other cell lines, uncontrolled expression may have unknown outcomes. To make gene therapy of hemophilia more resembling to the nature, we supposed f8 promoter might be a good candidate for driving downstream f8 coding sequence. Materials and Methods: To test our hypothesis, we designed and constructed a DNA construct by PCR, which harbors EGFP coding sequence downstream to mouse f8 promoter and transfected it to a mouse hepatoma cell line. Transfection was assayed qualitatively by fluorescent microscopy. Results: Fluorescence was detected in transfected cells a sign of presence of EGFP. Conclusion: f8 promoter can drive expression of downstream genes, a capability which and may have potential to be used in gene therapy of hemophilia. A conclusion that should be examined by further studies. [ABSTRACT FROM AUTHOR]

Interleukin- 18 (IL-18) has been reported to inhibit hepatitis B virus (HBV) replication in the liver of HBV transgenic mice; however, the molecular mechanism of its antiviral effect has not been fully understood. In the present study, it was shown that IL-18 and its receptors (IL- 18R) were constitutively expressed in hepatoma cell lines HepG2 and HepG2.2. 15 as well as normal liver cell line HL-7702. We demonstrated that IL-18 directly inhibited HBV replication in HepG2.2. 15 cells via downregulating the activities of HBV core and X gene promoters. The suppressed HBV replication by IL- 18 could be rescued by the administration of BAY1 1-7082, an inhibitor of transcription factor NF-κB. On the other hand, it was of interest that IL-I 8 promoted HepG2 cell metastasis and migration dose dependently in both wound-healing assays and Transwell assays. The underlying mechanism could be partially attributable to the increased activities of extracellular matrix metalloproteinase (MMP)-9, MMP-3, and MMP-2 by IL-18, which upregulated the mRNA levels of MMP-3 and MMP-9 in a NF-κB-dependent manner. Furthermore, it was confirmed that expression of IL-1811L-l8R and most MMPs were remarkably upregulated in hepatocellular carcinoma (HCC) liver cancer tissue specimens, suggesting that IL-1 8/IL-18R-triggered signaling pathway was closely related to HCC metastasis in vivo. Therefore, we revealed the dual effects of IL-18 in human hepatocytes: it not only inhibited HBV replication but also promoted hepatoma cells metastasis and migration. NF-κB played a critical role in both effects. Our work contributed to a deeper understanding of the biological function of IL-l8 in human hepatocytes. [ABSTRACT FROM AUTHOR]

Policosanol, a well-defined mixture of very long chain primary alcohols that is available as a nutraceutical product, has been reported to lower blood cholesterol levels. The present studies demonstrate that policosanol promotes the phosphorylation of AMP-kinase and HMG-CoA reductase in hepatoma cells and in mouse liver after intragastric administration, providing a possible means by which policosanol might lower blood cholesterol levels. Treatment of hepatoma cells with policosanol produced a 2.5-fold or greater increase in the phosphorylation of AMP-kinase and HMG-CoA reductase, and increased the phosphorylation of Ca/calmodulin-dependent kinase kinase (CaMKK), an upstream AMP-kinase kinase. Intragastric administration of policosanol to mice similarly increased the phosphorylation of hepatic HMG-CoA reductase and AMP-kinase by greater than 2-fold. siRNA-mediated suppression of fatty aldehyde dehydrogenase, fatty acyl-CoA synthetase 4, and acyl-CoA acetyltransferase expression in hepatoma cells prevented the phosphorylation of AMP-kinase and HMG-CoA reductase by policosanol, indicating that metabolism of these very long chain alcohols to activated fatty acids is necessary for the suppression of cholesterol synthesis, presumably by increasing cellular AMP levels. Subsequent peroxisomal β-oxidation probably augments this effect. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To demonstrate that a combination of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and Rana catesbeiana ribonuclease-6 (RC6) exerts synergistic cytotoxic effects on human hepatoma cells. Materials and Methods: Human hepatoma cells (J5 and HepG2) were treated with various concentrations of BCNU or RC6. The survival rate was determined by XTT assay. Apoptosis was determined by fluorescence-activated cell sorting analysis with propidium iodide/annexin-V double stain. Caspase activation was determined by Western blot assay. Results: BCNU and RC6 are able to inhibit the cell growth of hepatoma cells in a dose-dependent manner. BCNU combined with RC6 exerts a synergistic cytotoxic effect on hepatoma cells. Normal cells had less cytotoxicity than on hepatoma cells with BCNU/RC6 treatment. In addition, apoptosis was observed in hepatoma cells with BCNU treatment, RC6 treatment, and combination treatment. Our data also showed that combination treatment can activate the caspase-9/caspase-3 cascade obviously in hepatoma cells. Conclusion: Combination treatment with BCNU and RC6 exerts a synergistic cytotoxic effect on hepatoma cells. [Copyright &y& Elsevier]

Abstract: Followed by the results of our previous in vitro report (Food Chem. Toxicol., 2007), the efficacy of the soybean fermentation products containing live bacteria (SCB) was demonstrated using a syngeneic animal model. Murine HBV-related hepatoma ML-1 cells, derived from inbred animals and tumorigenic in BALB/c mice, were implanted subcutaneously to the flank of BALB/c mice on day 0. Three days after implantation, SCB (1.0 or 1.3ml/mouse/day) or vehicle (water) was orally administrated daily until day 60. The results indicate that SCB significantly reduced (P <0.05) the volumes and weights of tumors during the experimental periods. Examination using TUNEL staining on section of tumors revealed apoptotic phenomenon of nuclear DNA double-strand breaks in the groups of mice received SCB. Immunohistochemistry further revealed an autophagic LC3-II punctate pattern. Of note, SCB induced autophagy in the absence or presence of apoptosis, whereas, apoptosis was observed only in combination with autophagy. In vitro study using autophagy inhibitor indicated that the induction of autophagy promoted apoptosis. These data imply that the suppression in tumor volumes and tumor weights by oral administration of SCB was due to the induction of apoptotic and autophagic cell death, which suggests therapeutic potential of SCB on HBV-related HCC. [Copyright &y& Elsevier]

Abstract: Hyperthermia is a promising treatment for carcinoma cells. The thermal injuries of two hepatoma carcinoma cell lines with the identical cytological grade, HepG2 and Hep3B cell lines, were investigated systematically in the present study. The homemade heating stage was used to provide a constant temperature between 40 and 70°C for thermal treatment. When the cells were exposed to temperatures ranging from 40 to 45°C, Hep3B cells had a lower thermotolerance than the HepG2 cells; however, the survival rate of these two cell lines was still high. The differences in thermotolerance between HepG2 and Hep3B cells were more significant at the range of 50–55°C than those at lower-level temperatures of 40–45°C. Furthermore, the viability of the cells was less than 10% when they were exposed to a supraphysiological temperature of 60°C for 5min; these cell lines suffered from injury saturation under that thermal treatment. The statistical analysis also concluded that Hep3B cells are more susceptible to heat stress than are the HepG2 cells when subjected to the thermal treatment applied in this work, the exception being when thermal injury saturation occurred. The kinematic parameters of the activation energy and frequency factor for HepG2 and Hep3B cells were also quantitatively determined herein. The activation energies (ΔE) for HepG2 and Hep3B cells were 170.17 and 152.44kJ/mol, respectively. Furthermore, the frequency factors (A) for HepG2 and Hep3B cells were 4.11×1024 and 1.07×1022 s−1, respectively. [ABSTRACT FROM AUTHOR]