Next, we evaluated whether hepatocyte Nox proteins played a role in the increased detection of ROS with HCV. Huh7 cells were transfected with JFH1 RNA or mock-transfected and analyzed for Nox mRNA levels by qRT-PCR.7 Cells were also transfected with subgenomic JFH1 RNA for comparison. All seven Nox mRNAs could be detected in these

cells (Supporting Table 2). Most of all, we found that Nox4 mRNA began to be significantly elevated in the JFH1 cells at 48 hours, and the increase persisted at least to day 17, at which point the increase was more than 10-fold (Fig. 2A). In addition, Nox1 mRNA increased significantly with JFH1, and the increase persisted at least to days 14 to 17 (Fig. 2B; some data not shown). In contrast, Duox2 mRNA increased between 48 and 96 hours with JFH1, but this increase was not sustained (Fig. 2C). Nox2, Nox3, Nox5, and Duox1 mRNAs did not increase with JFH1 (data not shown). Subgenomic JFH1SgLuc RNA, which supports NVP-BGJ398 in vivo viral RNA genome replication without producing virus particles, replicated in these cells as expected (Supporting Fig. 3) but did not elevate Nox1, Nox4, or Duox2 mRNAs (Fig. 2C,D). Thus, Nox1 and Nox4 mRNAs showed prolonged elevation with genotype 2a HCV

in cell culture, and the structural genes of HCV and/or generation of infectious virions appeared to be necessary for the increases. HCV also increased p22phox, NOXA1, NOXO1, and p67phox mRNAs (Supporting Fig. 4). Next, Huh7 cells that were either transfected with JFH1 RNA or see more infected Selleck Apitolisib with a virus-containing cell culture medium from JFH1 RNA-transfected cells (Supporting Fig. 5) were analyzed for the levels of Nox1 and Nox4 proteins by western blotting. Nox1 and Nox4 proteins increased with HCV RNA transfection as well as infection (Fig. 3A,B,D). Higher molecular weight bands (>65 kDa) were also detected, particularly in the presence of HCV. Furthermore, Nox1 and Nox4 proteins were significantly elevated in HCV-infected human liver versus uninfected liver

samples (Fig. 3C). Therefore, Nox1 and Nox4 proteins were significantly elevated in vitro and during natural infection in vivo in the presence of HCV. To examine whether Nox1 and Nox4 played a role in the virus-induced ROS elevation, we used siRNAs to specifically knock down Nox1 and Nox4 gene expression in these cells. Nox1 siRNA decreased the Nox1 protein level to 27.3% ± 19.2% of the level of the controls transfected with nontargeting siRNAs at 72 hours (P < 0.05); Nox4 siRNA decreased the Nox4 protein level to 45.2% ± 12.3% of the level of the controls at 72 hours (P < 0.05; Fig. 4A). In addition, Nox1 and Nox4 siRNAs significantly decreased H2O2 and intracellular superoxide concentrations in the JFH1 cells (Fig. 4B,C). Nox1 and Nox4 siRNAs did not decrease other Nox mRNAs and selectively decreased the target protein without affecting Nox4 and Nox1 proteins, respectively (Supporting Fig. 6; some data not shown).

Obesity-induced weight gain, increase in fasting blood glucose and insulin levels, and augmented expression of gluconeogenic genes were restored to normal only 3 months after AAV treatment. Thus, CPT1A- and, to a greater extent, Fulvestrant CPT1AM-expressing mice were protected against obesity-induced weight gain, hepatic steatosis, diabetes, and obesity-induced insulin resistance. In addition, genetically

obese db/db mice that expressed CPT1AM showed reduced glucose and insulin levels and liver steatosis. Conclusion: A chronic increase in liver FAO improves the obese metabolic phenotype, which indicates that AAV-mediated CPT1A expression could be a potential molecular therapy for obesity and diabetes. (HEPATOLOGY 2011) Obesity is a major risk factor for disorders ranging from insulin resistance and type 2 diabetes (T2D) to hepatic

steatosis and cardiovascular disease. The incidence of obesity is increasing worldwide and a concerted effort is being made to understand its pathogenesis. Two main mechanisms have been proposed to explain obesity-induced insulin resistance: on the one hand the ectopic deposition of triacylglyceride (TAG) outside the adipose tissue,1 and on the other, Selleckchem Decitabine the heightened inflammatory state of the adipose tissue and liver.2 However, the ultimate cause of obesity is an energy imbalance between intake and expenditure, leading to the accumulation of excess nutrients in lipid deposits. Therefore, any strategy able to tilt the balance towards fatty-acid oxidation (FAO) could improve obesity-induced disorders. Malonyl-CoA, derived from glucose metabolism and the first intermediate in lipogenesis, regulates FAO by inhibiting carnitine palmitoyltransferase 1 (CPT1). This makes CPT1 the rate-limiting step in mitochondrial fatty-acid β-oxidation. Short-term genetic studies that increased FAO in liver showed a decrease in hepatic TAG content3 and insulin resistance in obese rodents.4, find more 5 However, to date there is no successful approach to chronically increase FAO and improve whole-animal obesity-induced insulin resistance in vivo. Here we achieved hepatic gene transfer of CPT1A (CPT1

liver isoform) to obese mice by injecting adeno-associated viruses (AAV) into the tail vein. This led to a nonimmunoreactive, long-term increase in lipid oxidation. We also used a mutant but active form of CPT1A (CPT1AM6), which is insensitive to malonyl-CoA and therefore leads to a permanent increase in the rate of FAO, independently of the glucose-derived malonyl-CoA levels. Our results show that an increase in hepatic FAO through AAV-mediated gene transfer of CPT1A and CPT1AM reduced obesity-induced hepatic steatosis, weight gain, inflammation, diabetes, and insulin resistance in mice consuming a high-fat diet (HFD). Furthermore, CPT1AM expression also reduced glucose and insulin levels, and liver steatosis in genetically obese db/db mice.

[5] Notably, from these same studies, anti-HCV prevalence estimates among U.S. detainees with a history of injection drug use were exceptionally high, ranging from 32.3% to 82.8%.[5] Given the large estimated number of detained persons worldwide and

the consistently high estimated prevalence among detainees in many countries where data are available, estimates of the anti-HCV burden that exclude detainees are likely underestimates. National, regional, and global estimates of anti-HCV prevalence in detainee populations are needed to Selleck Selumetinib produce better, “truer” estimates of the burden of HCV infection.[6] In this issue of Hepatology, Larney et al. provide regional and global estimates of anti-HCV prevalence among detainees in “prisons and other closed settings”.[7] Prisons and other closed settings was defined as prisons, BMS-907351 order jails, juvenile detention facilities, pretrial detention centers, and extrajudicial detention centers for people who use drugs and excluded psychiatric institutions and immigration detention

facilities. Estimates were based upon systematic review and meta-analysis of 93 studies reported between 1990 and September 2012. Specifically, regional summary prevalence estimates were produced using meta-analytic techniques, and, in turn, regional summary prevalence estimates were summarized using meta-analysis to produce a global summary prevalence estimate. To produce regional and global estimated counts

of anti-HCV-positive prisoners, click here regional summary prevalence estimates were applied to the number of prisoners reported or estimated in the region. Regional summary estimates were based on varying numbers of studies (from 1 in Central Asia to 39 in Western Europe) and showed considerable heterogeneity (I2 >94% in all regions). The global summary prevalence estimate for general detainees was 26% (95% confidence interval [CI]: 23%-29%) and for detainees with a history of injection drug use (k = 51) was 64% (95% CI: 58%-70%). The researchers estimated that 2.2 million (range, 1.4-2.9 million) detainees globally are anti-HCV positive. With this article, Larney et al. make a significant contribution to the literature. The search strategy and selection criteria for the review cast a broad, inclusive net, bringing together a large, international sample of anti-HCV prevalence studies among detainee populations. They identify and highlight national and regional differences in the availability of anti-HCV prevalence data from detainee populations, as well as variability of anti-HCV prevalence estimates from detainee populations within and across countries and regions. Perhaps most importantly, they demonstrate and underscore the problem of elevated prevalence of anti-HCV in detainee populations and begin to quantify the global scope of the problem at a critical time in history.

[5] Notably, from these same studies, anti-HCV prevalence estimates among U.S. detainees with a history of injection drug use were exceptionally high, ranging from 32.3% to 82.8%.[5] Given the large estimated number of detained persons worldwide and

the consistently high estimated prevalence among detainees in many countries where data are available, estimates of the anti-HCV burden that exclude detainees are likely underestimates. National, regional, and global estimates of anti-HCV prevalence in detainee populations are needed to learn more produce better, “truer” estimates of the burden of HCV infection.[6] In this issue of Hepatology, Larney et al. provide regional and global estimates of anti-HCV prevalence among detainees in “prisons and other closed settings”.[7] Prisons and other closed settings was defined as prisons, Stem Cells inhibitor jails, juvenile detention facilities, pretrial detention centers, and extrajudicial detention centers for people who use drugs and excluded psychiatric institutions and immigration detention

facilities. Estimates were based upon systematic review and meta-analysis of 93 studies reported between 1990 and September 2012. Specifically, regional summary prevalence estimates were produced using meta-analytic techniques, and, in turn, regional summary prevalence estimates were summarized using meta-analysis to produce a global summary prevalence estimate. To produce regional and global estimated counts

of anti-HCV-positive prisoners, selleckchem regional summary prevalence estimates were applied to the number of prisoners reported or estimated in the region. Regional summary estimates were based on varying numbers of studies (from 1 in Central Asia to 39 in Western Europe) and showed considerable heterogeneity (I2 >94% in all regions). The global summary prevalence estimate for general detainees was 26% (95% confidence interval [CI]: 23%-29%) and for detainees with a history of injection drug use (k = 51) was 64% (95% CI: 58%-70%). The researchers estimated that 2.2 million (range, 1.4-2.9 million) detainees globally are anti-HCV positive. With this article, Larney et al. make a significant contribution to the literature. The search strategy and selection criteria for the review cast a broad, inclusive net, bringing together a large, international sample of anti-HCV prevalence studies among detainee populations. They identify and highlight national and regional differences in the availability of anti-HCV prevalence data from detainee populations, as well as variability of anti-HCV prevalence estimates from detainee populations within and across countries and regions. Perhaps most importantly, they demonstrate and underscore the problem of elevated prevalence of anti-HCV in detainee populations and begin to quantify the global scope of the problem at a critical time in history.

[5] Notably, from these same studies, anti-HCV prevalence estimates among U.S. detainees with a history of injection drug use were exceptionally high, ranging from 32.3% to 82.8%.[5] Given the large estimated number of detained persons worldwide and

the consistently high estimated prevalence among detainees in many countries where data are available, estimates of the anti-HCV burden that exclude detainees are likely underestimates. National, regional, and global estimates of anti-HCV prevalence in detainee populations are needed to check details produce better, “truer” estimates of the burden of HCV infection.[6] In this issue of Hepatology, Larney et al. provide regional and global estimates of anti-HCV prevalence among detainees in “prisons and other closed settings”.[7] Prisons and other closed settings was defined as prisons, PXD101 in vitro jails, juvenile detention facilities, pretrial detention centers, and extrajudicial detention centers for people who use drugs and excluded psychiatric institutions and immigration detention

facilities. Estimates were based upon systematic review and meta-analysis of 93 studies reported between 1990 and September 2012. Specifically, regional summary prevalence estimates were produced using meta-analytic techniques, and, in turn, regional summary prevalence estimates were summarized using meta-analysis to produce a global summary prevalence estimate. To produce regional and global estimated counts

of anti-HCV-positive prisoners, selleckchem regional summary prevalence estimates were applied to the number of prisoners reported or estimated in the region. Regional summary estimates were based on varying numbers of studies (from 1 in Central Asia to 39 in Western Europe) and showed considerable heterogeneity (I2 >94% in all regions). The global summary prevalence estimate for general detainees was 26% (95% confidence interval [CI]: 23%-29%) and for detainees with a history of injection drug use (k = 51) was 64% (95% CI: 58%-70%). The researchers estimated that 2.2 million (range, 1.4-2.9 million) detainees globally are anti-HCV positive. With this article, Larney et al. make a significant contribution to the literature. The search strategy and selection criteria for the review cast a broad, inclusive net, bringing together a large, international sample of anti-HCV prevalence studies among detainee populations. They identify and highlight national and regional differences in the availability of anti-HCV prevalence data from detainee populations, as well as variability of anti-HCV prevalence estimates from detainee populations within and across countries and regions. Perhaps most importantly, they demonstrate and underscore the problem of elevated prevalence of anti-HCV in detainee populations and begin to quantify the global scope of the problem at a critical time in history.

Additional Supporting Information may be found in the online version of this article. “
“Tumor recurrence and metastases are the major obstacles to improving the prognosis of patients with hepatocellular carcinoma (HCC). To identify novel risk factors associated with HCC recurrence and metastases, we have established a panel of recurrence-associated selleck compound microRNAs (miRNAs) by comparing miRNA expression in recurrent and nonrecurrent human HCC tissue samples using microarrays (recurrence is defined as recurrent disease occurring within a 2-year time point of

the original treatment). Among the panel, expression of the miR-216a/217 cluster was consistently and significantly up-regulated in HCC tissue samples and cell lines associated with early tumor recurrence, find more poor disease-free survival, and an epithelial-mesenchymal transition (EMT) phenotype. Stable overexpression of miR-216a/217-induced EMT increased the stem-like cell population, migration, and metastatic ability of epithelial HCC cells. Phosphatase and tensin homolog (PTEN) and mothers against decapentaplegic homolog 7 (SMAD7) were subsequently identified as two functional targets of miR-216a/217, and both PTEN and SMAD7 were down-regulated in HCC. Ectopic expression of PTEN or SMAD7 partially rescued

miR-216a/217-mediated EMT, cell migration, and stem-like properties of HCC cells. Previously, SMAD7 was shown to be a transforming growth factor beta (TGF-β) type 1 receptor antagonist. Here, we further demonstrated that overexpression of miR-216a/217 acted as a positive feedback regulator for the TGF-β pathway and the canonical pathway involved in the activation of phosphoinositide 3-kinase/protein kinase K (PI3K/Akt) signaling in HCC cells. selleck Additionally, activation of the TGF-β- and PI3K/Akt-signaling pathways

in HCC cells resulted in an acquired resistance to sorafenib, whereas blocking activation of the TGF-β pathway overcame miR-216a/217-induced sorafenib resistance and prevented tumor metastases in HCC. Conclusion: Overexpression of miR-216a/217 activates the PI3K/Akt and TGF-β pathways by targeting PTEN and SMAD7, contributing to hepatocarcinogenesis and tumor recurrence in HCC. (Hepatology 2013;58:629–641) Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and the third-leading cause of deaths from cancer worldwide. Recurrent disease is one of the most serious challenges for managing patients with HCC.[1] Although hepatic resection is a well-accepted therapy for early-stage HCC, many patients develop tumor recurrence and this converts the situation to a dismal prognosis.[2] Coupled with the inherent high resistance of HCC to chemotherapeutic drugs, recurrent disease forms the main cause of death in long-term evaluations.

Additional Supporting Information may be found in the online version of this article. “
“Tumor recurrence and metastases are the major obstacles to improving the prognosis of patients with hepatocellular carcinoma (HCC). To identify novel risk factors associated with HCC recurrence and metastases, we have established a panel of recurrence-associated Ganetespib datasheet microRNAs (miRNAs) by comparing miRNA expression in recurrent and nonrecurrent human HCC tissue samples using microarrays (recurrence is defined as recurrent disease occurring within a 2-year time point of

the original treatment). Among the panel, expression of the miR-216a/217 cluster was consistently and significantly up-regulated in HCC tissue samples and cell lines associated with early tumor recurrence, MK-8669 research buy poor disease-free survival, and an epithelial-mesenchymal transition (EMT) phenotype. Stable overexpression of miR-216a/217-induced EMT increased the stem-like cell population, migration, and metastatic ability of epithelial HCC cells. Phosphatase and tensin homolog (PTEN) and mothers against decapentaplegic homolog 7 (SMAD7) were subsequently identified as two functional targets of miR-216a/217, and both PTEN and SMAD7 were down-regulated in HCC. Ectopic expression of PTEN or SMAD7 partially rescued

miR-216a/217-mediated EMT, cell migration, and stem-like properties of HCC cells. Previously, SMAD7 was shown to be a transforming growth factor beta (TGF-β) type 1 receptor antagonist. Here, we further demonstrated that overexpression of miR-216a/217 acted as a positive feedback regulator for the TGF-β pathway and the canonical pathway involved in the activation of phosphoinositide 3-kinase/protein kinase K (PI3K/Akt) signaling in HCC cells. this website Additionally, activation of the TGF-β- and PI3K/Akt-signaling pathways

in HCC cells resulted in an acquired resistance to sorafenib, whereas blocking activation of the TGF-β pathway overcame miR-216a/217-induced sorafenib resistance and prevented tumor metastases in HCC. Conclusion: Overexpression of miR-216a/217 activates the PI3K/Akt and TGF-β pathways by targeting PTEN and SMAD7, contributing to hepatocarcinogenesis and tumor recurrence in HCC. (Hepatology 2013;58:629–641) Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and the third-leading cause of deaths from cancer worldwide. Recurrent disease is one of the most serious challenges for managing patients with HCC.[1] Although hepatic resection is a well-accepted therapy for early-stage HCC, many patients develop tumor recurrence and this converts the situation to a dismal prognosis.[2] Coupled with the inherent high resistance of HCC to chemotherapeutic drugs, recurrent disease forms the main cause of death in long-term evaluations.

7C). Because UCP2 is known to be up-regulated in a tissue-dependent manner during fasting and can regulate insulin secretion,19 mRNA levels of UCP2 were measured in adipose and liver tissue. In the fed state, UCP2 mRNA was significantly higher in Hint2−/− than in Hint2+/+ WAT (Fig. 5E). After fasting, UCP2 MK-2206 manufacturer increased in Hint2+/+ WAT but did not increase further in Hint2−/− WAT. In the liver, UCP2 was similar in both fed groups and decreased after fasting in Hint2+/+ (Fig. 5E). To test for expression of Hint2 in adipose tissue, immunoblotting was performed

in WAT and brown adipose tissue (BAT). Hint2 was only detected in BAT (Fig. 5F). No differences were detected between mitochondria www.selleckchem.com/products/apo866-fk866.html from Hint2−/− and Hint2+/+ livers in the expression of respiratory complexes (Supporting Fig. 4) and in the individual activities of complexes I, II, and III (Fig. 6A). However, the activity of the linked complex II-III was reduced by 60% in Hint2−/− mitochondria. Accordingly, succinate-linked state 3 respiration was decreased by 44%, and pyruvate-linked respiration was decreased by 35% in Hint2−/− mice (Fig. 6B). The content of total coenzyme Q was lower in Hint2−/− livers than in Hint2+/+ livers (Fig. 4C). To determine whether a change in the expression of genes involved in the biosynthesis of coenzyme Q was responsible, the mRNA expression of polyisoprenyl

diphosphate synthases 1 and 2, Coq2, Coq3, Coq4, Coq5, Coq6, Coq7, Coq8, and Coq9 was compared in Hint2−/− and Hint2+/+ livers via real-time PCR. Only Coq8 increased 2.5-fold in Hint2−/− at 20 weeks and Coq9 increased 1.6-fold at 10 weeks (data not shown). To confirm the link between HINT2 expression and the altered energy metabolism, we generated HepG2 cell lines that expressed varying levels of HINT2 (Supporting Fig. 5). The activities

of the individual selleck chemical respiratory chain complexes I, II, III, and IV were not different among the HepG2 variants (data not shown). The silencing of HINT2 (HepG2-siRNA-HINT2) was associated with a 30% decrease in state 3 respiration in the presence of pyruvate and succinate, whereas an overexpression of HINT2 did not influence the state 3 respiration (Fig. 6D). When expressed relative to citrate synthase, oxygen consumption in HepG2-siRNA-HINT2 cells was reduced (Fig. 6E). No differences in state 4 respiration were observed between the cell lines (data not shown). Hint2−/− hepatocytes produced a 1.5-fold higher level of reactive oxygen species (Supporting Fig. 6A). Activation of hypoxia-inducible transcription factor (Hif) signaling was examined via immunohistochemistry. Activation of Hif-2α but not Hif-1α was higher in Hint2−/− than in Hint2+/+ livers (Supporting Fig. 6B). To confirm the link between GDH activity and the absence of Hint2, enzymatic assays were repeated in lysates of HepG2 over- and under-expressing cells.

7C). Because UCP2 is known to be up-regulated in a tissue-dependent manner during fasting and can regulate insulin secretion,19 mRNA levels of UCP2 were measured in adipose and liver tissue. In the fed state, UCP2 mRNA was significantly higher in Hint2−/− than in Hint2+/+ WAT (Fig. 5E). After fasting, UCP2 Anti-infection Compound Library mw increased in Hint2+/+ WAT but did not increase further in Hint2−/− WAT. In the liver, UCP2 was similar in both fed groups and decreased after fasting in Hint2+/+ (Fig. 5E). To test for expression of Hint2 in adipose tissue, immunoblotting was performed

in WAT and brown adipose tissue (BAT). Hint2 was only detected in BAT (Fig. 5F). No differences were detected between mitochondria Selleckchem Sunitinib from Hint2−/− and Hint2+/+ livers in the expression of respiratory complexes (Supporting Fig. 4) and in the individual activities of complexes I, II, and III (Fig. 6A). However, the activity of the linked complex II-III was reduced by 60% in Hint2−/− mitochondria. Accordingly, succinate-linked state 3 respiration was decreased by 44%, and pyruvate-linked respiration was decreased by 35% in Hint2−/− mice (Fig. 6B). The content of total coenzyme Q was lower in Hint2−/− livers than in Hint2+/+ livers (Fig. 4C). To determine whether a change in the expression of genes involved in the biosynthesis of coenzyme Q was responsible, the mRNA expression of polyisoprenyl

diphosphate synthases 1 and 2, Coq2, Coq3, Coq4, Coq5, Coq6, Coq7, Coq8, and Coq9 was compared in Hint2−/− and Hint2+/+ livers via real-time PCR. Only Coq8 increased 2.5-fold in Hint2−/− at 20 weeks and Coq9 increased 1.6-fold at 10 weeks (data not shown). To confirm the link between HINT2 expression and the altered energy metabolism, we generated HepG2 cell lines that expressed varying levels of HINT2 (Supporting Fig. 5). The activities

of the individual find more respiratory chain complexes I, II, III, and IV were not different among the HepG2 variants (data not shown). The silencing of HINT2 (HepG2-siRNA-HINT2) was associated with a 30% decrease in state 3 respiration in the presence of pyruvate and succinate, whereas an overexpression of HINT2 did not influence the state 3 respiration (Fig. 6D). When expressed relative to citrate synthase, oxygen consumption in HepG2-siRNA-HINT2 cells was reduced (Fig. 6E). No differences in state 4 respiration were observed between the cell lines (data not shown). Hint2−/− hepatocytes produced a 1.5-fold higher level of reactive oxygen species (Supporting Fig. 6A). Activation of hypoxia-inducible transcription factor (Hif) signaling was examined via immunohistochemistry. Activation of Hif-2α but not Hif-1α was higher in Hint2−/− than in Hint2+/+ livers (Supporting Fig. 6B). To confirm the link between GDH activity and the absence of Hint2, enzymatic assays were repeated in lysates of HepG2 over- and under-expressing cells.

Our results indicate that HBx does not seem to cooperate with constitutively active NRAS

to induce liver tumorigenesis in HBx/NRAS animals. This was evident from the relatively low ALT levels in serum (Table 1), the low tumor multiplicity, and the liver weight PD0325901 datasheet to whole mass percentage (Fig. 3A,B) in HBx/NRAS mice. HBx has been suggested to up-regulate the Ras signaling pathway.17 Perhaps HBx expression and activated Ras are redundant in this transformation assay. Even when all the transgenes were coinjected (HBx/NRAS/shp53), there was only a marginally significant increase (P < 0.05) in tumorigenicity in comparison with HBx alone. Moreover, no significant increase in tumorigenicity was seen in HBx/NRAS animals versus animals with NRAS alone (Fig. 3A). Our results indicate that HBx up-regulates the Wnt signaling pathway, and this may play a role in liver tumorigenesis (Fig. 4), whereas constitutively active NRAS seems to induce hyperplasia, probably via the

RAF/mitogen-activated protein kinase kinase/extracellular signal-regulated kinase and/or phosphoinositide 3-kinase (PI3K)/v-akt murine thymoma PARP assay viral oncogene homolog 1 (AKT) associated pathways (Fig. 5). In addition, we detected high levels of CD45-positive staining cells in livers of animals injected with HBx alone or in combination with other transgenes (Supporting Information Fig. 4). These cells could represent infiltrating lymphocytes, which are often associated with HBV infection. Indeed, the HBx protein would be predicted to act as a foreign antigen in our system, unlike previously reported HBx transgenic mouse models. HBx-induced inflammation may play some this website role in HCC progression; this hypothesis could be tested

with our model. Elevated pAkt levels were detected by IHC in experimental animals injected with NRAS alone or in combination with shp53, and this indicates that NRAS is likely signaling via the Pi3k/Akt pathway (Fig. 5). HBx has been previously shown to activate the WNT/CTNNB1 signaling pathway in human hepatoma cell lines.8HBx antigen has also been associated with the accumulation of CTNNB1 in the cytoplasm and/or nucleus and the up-regulation of the HBx antigen effector up-regulated gene 11, which results in increased activation of CTNNB1.18 The CTNNB1 staining pattern can be correlated with the histopathological types of liver tumors.19 The absence of nuclear staining and strong membranous staining with rare, weak cytoplasmic expression of Ctnnb1 suggested that the hyperplastic nodules induced by HBx or HBx/shp53 were adenocarcinomas or poorly differentiated HCC (Fig. 4). We did not detect any activation of Stat3 in liver tumors expressing HBx by IHC in our experimental cohorts with a phospho-Stat3 (Tyr705)–specific antibody (data not shown) despite the previous suggestion that HBx activates Stat3.