• Biomarker
• Hepatitis B surface antigen
• Hepatocellular carcinoma
• Oncoprotein
• Tumorigenesis

• 82073090 National Natural Science Foundation of China
• 202303021211220 Shanxi Province "136" Revitalization Medical Project Construction Funds, Fundamental Research Program of Shanxi Province - Free Exploration Category
• 2023M741275 China Postdoctoral Science Foundation
• 2024ZHA NCHI07 Research and Innovation Team Project for Scientific Breakthroughs at Shanxi Bethune Hospital
• Shanxi Province “136” Revitalization Medical Project Construction Funds, Fundamental Research Program of Shanxi Province - Free Exploration Category

• hepatitis B virus
• hepatocellular carcinoma
• molecular mechanism
• pre-S mutant
• therapeutic target

• Humans
• Carcinoma, Hepatocellular/virology
• Carcinoma, Hepatocellular/pathology
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/drug therapy
• Liver Neoplasms/virology
• Liver Neoplasms/pathology
• Liver Neoplasms/drug therapy
• Liver Neoplasms/genetics
• Hepatitis B virus/genetics
• Hepatitis B virus/pathogenicity
• Mutation
• Carcinogenesis/genetics
• Hepatitis B Surface Antigens/genetics
• Hepatitis B, Chronic/complications
• Hepatitis B, Chronic/virology

• China Medical University, Taichung, Taiwan

• AAV vectors
• VEGF
• VEGF peptides
• angiogenesis
• bevacizumab
• oncolytic viruses
• tumor viruses
• viral capsids

• Humans
• Vascular Endothelial Growth Factor A/metabolism
• Animals
• Viruses/metabolism
• Genetic Vectors/metabolism
• Neovascularization, Pathologic
• Neoplasms/therapy
• Neoplasms/virology
• Neoplasms/pathology
• Neoplasms/metabolism
• Genetic Therapy/methods

• PID2022-141799OB-I00 Spanish Ministerio de Ciencia, Investigación y Universidades. AEI

• Evolution
• Hepatitis B virus
• Hepatocellular carcinoma
• Quasispecies

• Hepatitis B virus/genetics
• Hepatitis B virus/classification
• Humans
• Genome, Viral
• Genetic Variation
• Evolution, Molecular
• Hepatitis B/virology
• Genotype
• Antiviral Agents/therapeutic use
• Antiviral Agents/pharmacology

• Bibliometrics
• Endoplasmic reticulum
• Hepatocellular carcinoma
• Research trends
• Stress

• 23JRRA0975 Natural Science Foundation of Gansu Province

• HBsAg isoforms
• diagnosis
• hepatitis B virus (HBV)
• large surface protein (LHB)
• middle surface protein (MHB)
• mutation
• new antivirals
• new biomarkers
• small surface protein (SHB)
• vaccine

• Humans
• Hepatitis B Surface Antigens/blood
• Hepatitis B Surface Antigens/immunology
• Hepatitis B Surface Antigens/metabolism
• Protein Isoforms
• Hepatitis B virus/genetics
• Hepatitis B virus/immunology
• Antiviral Agents/therapeutic use
• Hepatitis B/diagnosis
• Hepatitis B/immunology
• Hepatitis B/prevention & control
• Biomarkers/blood
• Biomarkers/metabolism
• Hepatitis B, Chronic/drug therapy
• Hepatitis B, Chronic/diagnosis
• Hepatitis B, Chronic/immunology
• Hepatitis B Vaccines/immunology
• Hepatitis B Vaccines/administration & dosage

• 200110 Wellcome Trust

• EBV
• HBV
• HCV
• HIV
• HPV
• HTLV
• KSHV

• Humans
• Epstein-Barr Virus Infections/complications
• Herpesvirus 4, Human
• Herpesvirus 8, Human/genetics
• Neoplasms/etiology
• Vascular Endothelial Growth Factor A/genetics
• Virus Diseases/complications

• 1402-2-75-26034 Iran University of Medical Sciences
• Research Department of the School of Medicine Shahid Beheshti University of Medical Sciences

• GGH
• HBV
• iDNA
• long-read sequencing
• pre-S
• tumor evolution

• Humans
• Carcinoma, Hepatocellular/genetics
• Hepatitis B virus/genetics
• DNA, Viral/genetics
• Liver Neoplasms/genetics
• Hepatocytes
• Mutation
• Antiviral Agents/therapeutic use

• R01 CA125642 NCI NIH HHS
• R01 CA202769 NCI NIH HHS
• R44 CA165312 NCI NIH HHS
• R56 AI179574 NIAID NIH HHS

• FIH-1
• HBV-miR-3
• HBV-related HCC
• angiogenesis

• National Key Research and Development Program of China
• project for disciplines of excellence, West China Hospital, Sichuan University

• Angiogenesis
• Early detection
• Liver cancer
• Marker
• Metastasis
• MicroRNA

• Humans
• MicroRNAs/genetics
• MicroRNAs/metabolism
• Liver Neoplasms/genetics
• Neovascularization, Pathologic/genetics
• Signal Transduction
• Biomarkers, Tumor/genetics
• Biomarkers, Tumor/metabolism
• Gene Expression Regulation, Neoplastic
• Tumor Microenvironment

• hepatitis B virus
• hepatocellular carcinoma
• monocyte chemoattractant protein-1
• pre-S2 gene deletion mutation
• recurrence

• Humans
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/surgery
• Chemokine CCL2/genetics
• Chemokine CCL2/metabolism
• Gene Deletion
• Hepatitis B
• Hepatitis B Surface Antigens/genetics
• Hepatitis B Surface Antigens/metabolism
• Hepatitis B virus/genetics
• Hepatitis B, Chronic/complications
• Hepatitis B, Chronic/genetics
• Hepatitis B, Chronic/pathology
• Liver Neoplasms/genetics
• Liver Neoplasms/surgery
• Liver Neoplasms/pathology
• Mutation

Viral mutations at the preS region of hepatitis B virus (HBV) significantly increase the risk of developing hepatocellular carcinoma (HCC). Compared to HBV preS deletion, the oncogenic effect of preS combo mutation has rarely been investigated. With a cohort including 2114 subjects, we demonstrated that preS combo mutations G2950A/G2951A/A2962G/C2964A and C3116T/T31C significantly increased the risk of HCC in patients without antiviral treatment, whereas preS2 deletion significantly increased the risk of HCC in patients with antiviral treatment. The prevalence of C3116T/T31C (43.61%) was higher than preS2 deletion (7.16%). By using Sleeping Beauty mouse models and in vitro experiments, we found G2950A/G2951A/A2962G/C2964A, C3116T/T31C, and preS2 deletion promoted hepatocarcinogenesis by increasing levels of inflammatory cytokines, activating STAT3 pathway, enhancing endoplasmic reticulum stress, and altering gene expression profiles in inflammation- and metabolism-related pathways. These results suggest that preS combo mutations G2950A/G2951A/A2962G/C2964A and C3116T/T31C had similar oncogenic effects of preS2 deletion and should also be monitored.

This study aimed to elucidate the effects and underlying mechanisms of hepatitis B virus (HBV) preS mutations on hepatocarcinogenesis. The effect of the preS mutations on hepatocellular carcinoma (HCC) occurrence was evaluated using a prospective cohort study with 2114 HBV-infected patients, of whom 612 received antiviral treatments. The oncogenic functions of HBV preS mutations were investigated using cancer cell lines and Sleeping Beauty (SB) mouse models. RNA-sequencing and microarray were applied to identify key molecules involved in the mutant-induced carcinogenesis. Combo mutations G2950A/G2951A/A2962G/C2964A and C3116T/T31C significantly increased HCC risk in patients without antiviral treatment, whereas the preS2 deletion significantly increased HCC risk in patients with antiviral treatment. In SB mice, the preS1/preS2/S mutants induced a higher rate of tumor and higher serum levels of inflammatory cytokines than did wild-type counterpart. The preS1/preS2/S mutants induced altered gene expression profiles in the inflammation- and metabolism-related pathways, activated pathways of endoplasmic reticulum (ER) stress, affected the response to hypoxia, and upregulated the protein level of STAT3. Inhibiting the STAT3 pathway attenuated the effects of the preS1/preS2/S mutants on cell proliferation. G2950A/G2951A/A2962G/C2964A, C3116T/T31C, and preS2 deletion promote hepatocarcinogenesis via inducing ER stress, metabolism alteration, and STAT3 pathways, which might be translated into HCC prophylaxis.

Although surgical resection is available as a potentially curative therapy for hepatocellular carcinoma (HCC), high recurrence of HCC after surgery remains a serious obstacle for long-term patient survival. Therefore, the discovery of valuable prognostic biomarkers for HCC recurrence is urgently needed. Pre-S2 mutant is a mutant form of hepatitis B virus (HBV) large surface protein which is expressed from the HBV surface gene harboring deletion mutations spanning the pre-S2 gene segment. Pre-S2 mutant-positive HCC patients have been regarded as a high-risk population of HCC recurrence after resection surgery and display increased immune checkpoint programmed death ligand 1 (PD-L1) expression and pro-tumor regulatory T cells (Tregs) infiltration in tumor tissues. In this study, the association of higher levels of PD-L1 expression and Tregs infiltration in tumor tissues with post-operative HCC recurrence in pre-S2 mutant-positive HCC patients was evaluated. We found that patients with pre-S2 mutant in combination with higher levels of PD-L1 expression and Tregs infiltration in tumor tissues were independently associated with a higher risk of HCC recurrence (hazard ratio, 4.109; p value = 0.0011) and poorer recurrence-free survival (median, 8.2 versus 18.0 months; p value = 0.0004) than those of patients with either one or two of these three biomarkers. Furthermore, a combination of pre-S2 mutant, intra-tumoral PD-L1 expression, and tumor-infiltrating Tregs exhibited superior performance in identifying patients at a higher risk of HCC recurrence (area under the receiver operating characteristic curve, 0.8400). Collectively, this study suggests that higher levels of PD-L1 expression and Tregs infiltration in tumor tissues predicted a higher risk of HCC recurrence in pre-S2 mutant-positive HCC patients after curative surgical resection.

• curative surgical resection
• hepatitis B virus
• hepatocellular carcinoma
• pre-S2 mutant
• programmed death ligand 1
• recurrence
• regulatory T cells

Chronic hepatitis B (CHB) virus infection is the most important cause of HCC and is also associated with tumor progression. The development of viral biomarkers for HCC prognosis is critical in evaluating relative risks to recurrence in the CHB HCC patients. We report that the large HBV surface protein (LHBS) expression increased in the tumors, implicating that it played a significant role in tumor development. To detect the LHBS in serum and evaluate its association with HCC progression, we developed a sandwich ELISA method for LHBS. The mouse monoclonal antibodies for the pre-S1, pre-S2, and HBS regions were in-house generated and constructed into a chemiluminescent sandwich ELISA system, which allowed sensitive and quantitative measurement of the protein. Using this ELISA assay, we estimated the expression of LHBS in CHB and HCC patients. We found that the serum LHBS level was correlated with the HBS but not the viral titer in serum, indicating that HBV surface proteins’ expression does not mainly depend on viral replication. Moreover, both serum LHBS and HBS levels were lower in the HCC patients than in the CHB. The liver LHBS signals, detected by immunohistochemical staining, showed significant correlations with the serum LHBS and HBS levels. In addition, the more elevated serum LHBS but not HBS level was significantly associated with cirrhosis and worse disease-free and overall survival rates, based on the multivariate analysis. Conclusion: LHBS plays a specific role in tumor progression and is an independent parameter associated with HCC recurrence. Serum LHBS represents a novel noninvasive biomarker for HCC patients with a worse prognosis after surgery.

• ELISA
• HBV surface antigen
• chronic hepatitis B
• hepatocellular carcinoma
• prognosis

• Animals
• Biomarkers
• Carcinoma, Hepatocellular/surgery
• Hepatitis B virus
• Humans
• Liver Neoplasms/surgery
• Membrane Proteins
• Mice

With the advent of combination antiretroviral therapy (cART), overall survival has been improved, and the incidence of acquired immunodeficiency syndrome (AIDS)-defining cancers has also been remarkably reduced. However, non-AIDS-defining cancers among human immunodeficiency virus-1 (HIV-1)-associated malignancies have increased significantly so that cancer is the leading cause of death in people living with HIV in certain highly developed countries, such as France. However, it is currently unknown how HIV-1 infection raises oncogenic virus-mediated cancer risks in the HIV-1 and oncogenic virus co-infected patients, and thus elucidation of the molecular mechanisms for how HIV-1 expedites the oncogenic viruses-triggered tumorigenesis in the co-infected hosts is imperative for developing therapeutics to cure or impede the carcinogenesis. Hence, this review is focused on HIV-1 and oncogenic virus co-infection-mediated molecular processes in the acceleration of non-AIDS-defining cancers.

• F31 DA053151 NIDA NIH HHS
• F31DA053151 NIDA NIH HHS

Hepatitis B virus (HBV) infection causes acute and chronic liver diseases, including severe hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). Interferon alpha 2a (IFNα-2a) is commonly used for treating chronic HBV infection. However, its efficacy remains relatively low. Yet, the immunological and molecular mechanisms for successful IFNα-2a treatment remain elusive. One issue is whether the application of increasing IFNα doses may modulate cellular processes and HBV replication in hepatic cells. In the present study, we focused on the interaction of IFNα signaling with other cellular signaling pathways and the consequence for HBV replication. The results showed that with the concentration of 6000 U/ml IFNα-2a treatment downregulated the activity of not only the Akt/mTOR signaling but also the AMPK signaling. Additionally, IFNα-2a treatment increased the formation of the autophagosomes by blocking autophagic degradation. Furthermore, IFNα-2a treatment inhibited the Akt/mTOR signaling and initiated autophagy under low and high glucose concentrations. In reverse, inhibition of autophagy using 3-methyladenine (3-MA) and glucose concentrations influenced the expression of IFNα-2a-induced ISG15 and IFITM1. Despite of ISGs induction, HBV replication and gene expression in HepG2.2.15 cells, a cell model with continuous HBV replication, were slightly increased at high doses of IFNα-2a. In conclusion, our study indicates that IFNα-2a treatment may interfere with multiple intracellular signaling pathways, facilitate autophagy initiation, and block autophagic degradation, thereby resulting in slightly enhanced HBV replication.

• Autophagy
• Hepatitis B/drug therapy
• Hepatitis B virus/drug effects
• Hepatitis B virus/physiology
• Humans
• Interferon-alpha/pharmacology
• Virus Replication/drug effects

Mammalian target of rapamycin (mTOR) is a conserved Ser/Thr kinase that includes mTOR complex (mTORC) 1 and mTORC2. The mTOR pathway is activated in viral hepatitis, including hepatitis B virus (HBV) infection-induced hepatitis. Currently, chronic HBV infection remains one of the most serious public health issues worldwide. The unavailability of effective therapeutic strategies for HBV suggests that clarification of the pathogenesis of HBV infection is urgently required. Increasing evidence has shown that HBV infection can activate the mTOR pathway, indicating that HBV utilizes or hijacks the mTOR pathway to benefit its own replication. Therefore, the mTOR signaling pathway might be a crucial target for controlling HBV infection. Here, we summarize and discuss the latest findings from model biology research regarding the interaction between the mTOR signaling pathway and HBV replication.

• Autophagy
• Hepatitis B virus (HBV)
• Mammalian target of rapamycin (mTOR)
• Metabolism

Background: Hepatitis B virus (HBV) variants in the preS region have been associated with hepatocellular carcinoma (HCC). However, the effect of the preS variants on HCC prognosis remains largely unknown. We aimed to identify the preS variants that reliably predict postoperative prognosis in HCC.

Methods: RNA-seq data of 203 HCC patients retrieved from public database were screened for the preS variants related to HCC prognosis. The variants in the sera and tumors were then validated in our prospective cohort enrolling 103 HBV-associated HCC patients.

Results: By analyzing prognosis-related gene sets in the RNA-seq data, 12 HBV preS variants were associated with HCC recurrence. Of those, G40C and C147T in the sera predicted an unfavorable recurrence-free survival in our cohort (hazard ratio [HR]=2.18, 95% confidence interval [CI]=1.37-3.47, p=0.001 for G40C; HR=1.84, 95% CI=1.15-2.92, p=0.012 for C147T). G40C and C147T were significantly associated with microscopic vascular invasion, larger tumor size, and abnormal liver function. Multivariate Cox regression analysis showed that G40C significantly increased the risk of HCC recurrence in patients with postoperative antiviral treatment. The HCC prognosis-prediction model consisting of α-fetoprotein and G40C in the sera achieved the best performance (sensitivity=0.80, specificity=0.70, and area under the curve=0.79). Functional analysis indicated that these two variants were associated with cell proliferation, chromosome instability, carcinogenesis, metastasis, and anticancer drug resistance.

Conclusions: G40C and C147T are serological biomarkers for HCC prognosis and the prognostic model consisting of serological α-fetoprotein and G40C achieved the best performance in predicting postoperative prognosis.

• hepatitis B virus
• hepatocellular carcinoma
• prediction model
• prognosis
• viral variant

In traditional Chinese medicine (TCM), TCM syndrome is a key guideline, and Chinese materia medicas are widely used to treat hepatitis B virus- (HBV-) related hepatocellular carcinoma (HCC) according to different TCM syndromes. However, the prognostic value of TCM syndromes in HBV-related HCC patients has never been studied.

A retrospective cohort of HBV-related HCC patients at Shenzhen Traditional Chinese Medicine Hospital from December 2005 to October 2017 was analyzed. The prognostic value of TCM syndromes in HBV-related HCC patients was assessed by Kaplan–Meier survival curves and Cox analysis, and the TCM syndrome with the best prognosis of HBV-related HCC patients was determined. To further study the relevant mechanisms, key Chinese materia medicas (KCMMs) for the TCM syndrome with the best prognosis were summarized, and network pharmacology was also performed.

A total of 207 HBV-related HCC patients were included in this research, and we found that HBV-related HCC patients with TCM excess syndrome had better OS. Then, a total of eight KCMMs for TCM excess syndrome were identified, whose crucial ingredients included quercetin, beta-sitosterol, kaempferol, luteolin, and XH-14, and KCMMs could play a therapeutic role through MAPK, JAK-STAT, Wnt, Hippo, and other pathways. Moreover, TP53, SRC, STAT3, MAPK3, PIK3R1, HRAS, VEGFA, HSP90AA1, EGFR, and JAK2 were determined as the key targets.

We propose a new research method of “prognosis of TCM syndromes-KCMMs-network pharmacology” to reveal the prognostic value of TCM syndromes and the potential mechanism by which TCM syndromes affect prognosis.

Hepatitis B virus (HBV) infection is the major risk factor for hepatocellular carcinoma (HCC). Understanding the unique features for HBV-induced HCC can shed new light on the unmet needs in its early diagnosis and effective therapy. During decades of chronic hepatitis B, hepatocytes undergoing repeated damage and regeneration accumulate genetic changes predisposing to HCC development. In addition to traditional mutations in viral and cellular oncogenes, HBV integration into the cell chromosomes is an alternative genetic change contributing to hepatocarcinogenesis. A striking male dominance in HBV-related HCC further highlights an interaction between androgen sex hormone and viral factors, which contributes to the gender difference via stimulating viral replication and activation of oncogenes preferentially in male patients. Meanwhile, a novel circulating tumor biomarker generated by HBV integration shows great potential for the early diagnosis of HCC. These unique HBV-induced hepatocarcinogenic mechanisms provide new insights for the future development of superior diagnosis and treatment strategies.

Hepatitis B virus (HBV) infection is one of the important risk factors for hepatocellular carcinoma (HCC) worldwide, accounting for around 50% of cases. Chronic hepatitis B infection generates an inflammatory microenvironment, in which hepatocytes undergoing repeated cycles of damage and regeneration accumulate genetic mutations predisposing them to cancer. A striking male dominance in HBV-related HCC highlights the influence of sex hormones which interact with viral factors to influence carcinogenesis. HBV is also considered an oncogenic virus since its X and surface mutant proteins showed tumorigenic activity in mouse models. The other unique mechanism is the insertional mutagenesis by integration of HBV genome into hepatocyte chromosomes to activate oncogenes. HCC survival largely depends on tumor stages at diagnosis and effective treatment. However, early diagnosis by the conventional protein biomarkers achieves limited success. A new biomarker, the circulating virus–host chimera DNA from HBV integration sites in HCC, provides a liquid biopsy approach for monitoring the tumor load in the majority of HBV–HCC patients. To maximize the efficacy of new immunotherapies or molecular target therapies, it requires better classification of HCC based on the tumor microenvironment and specific carcinogenic pathways. An in-depth study may benefit both the diagnosis and treatment of HBV-related HCC.

• circulating tumor DNA
• hepatitis B virus (HBV)
• hepatocellular carcinoma (HCC)
• inflammation
• sex hormone
• virus integration

• biomarkers
• hepatitis B virus
• hepatocellular carcinoma
• pre-S deleted proteins
• pre-S gene deletions
• targets

• Animals
• Biomarkers
• Carcinoma, Hepatocellular/etiology
• Carcinoma, Hepatocellular/pathology
• Cell Cycle
• Cell Transformation, Viral
• Centrosome
• DNA Damage
• Disease Models, Animal
• Disease Susceptibility
• Endoplasmic Reticulum Stress
• Gene Deletion
• Gene Expression Regulation, Viral
• Hepatitis B/complications
• Hepatitis B/virology
• Hepatitis B Surface Antigens/genetics
• Hepatitis B virus/genetics
• Hepatitis B virus/physiology
• Humans
• Liver Cirrhosis/etiology
• Liver Cirrhosis/metabolism
• Liver Cirrhosis/pathology
• Liver Neoplasms/etiology
• Liver Neoplasms/pathology
• Prognosis
• Signal Transduction
• Virus Replication

Chronic hepatitis B virus (HBV) infection is a major risk factor for hepatocellular carcinoma (HCC), a leading cause of cancer-related death worldwide. The HCC patients who harbor HBV pre-S2 mutant, an oncoprotein that plays key roles in HCC development, have been closely associated with a worse prognosis after curative surgical resection, suggesting an urgent need for alternative therapeutic options to improve their survival. In this study, we aimed to evaluate the expression profiles of programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1), two of the most well-studied immune checkpoint molecules that promote tumor immune evasion, in tumor of the pre-S2 mutant-positive/high HCC patients.

We classified 40 HBV-related HCC patients into the pre-S2-positive/high and -negative/low groups by a next-generation sequencing-based approach. The fluorescent immunohistochemistry staining was performed to detect the expression of PD-1 and PD-L1 in HCC tissues of patients.

We showed that patients with either deletion spanning pre-S2 gene segment or high percentage of pre-S2 plus pre-S1+pre-S2 deletion (the pre-S2 mutant-positive/high group) exhibited a significantly higher density of PD-L1-positive cells in HCC tissues than those without. Moreover, the percentage of pre-S2 plus pre-S1+pre-S2 deletion displayed a high positive correlation with the density of PD-L1-positive cells in HCC tissues.

The increased expression of PD-L1 in tumor tissues of the pre-S2 mutant-positive HCC patients suggest that pre-S2 mutant may play a potential role in dysregulation of tumor immune microenvironment in the progression of HBV-related HCC, implicating for the development of future therapeutic strategies.

• hepatitis B virus
• hepatocellular carcinoma
• pre-S2 mutant
• programmed death 1
• programmed death ligand 1

• HBs
• HBx
• autophagosome
• autophagy
• hepatitis B virus
• viral replication

• Animals
• Antiviral Agents/pharmacology
• Antiviral Agents/therapeutic use
• Apoptosis
• Autophagosomes/metabolism
• Autophagy/drug effects
• Hepatitis B virus/metabolism
• Hepatitis B, Chronic/drug therapy
• Hepatitis B, Chronic/metabolism
• Hepatitis B, Chronic/virology
• Host Microbial Interactions
• Humans
• Lysosomes/metabolism
• Mice
• Trans-Activators/metabolism
• Viral Regulatory and Accessory Proteins/metabolism
• Virion/metabolism
• Virus Replication

Hepatitis B is the most prevalent viral hepatitis worldwide, affecting approximately one-third of the world’s population. Among HBV factors, the surface protein is the most sensitive biomarker for viral infection, given that it is expressed at high levels in all viral infection phases. The large HBV surface protein (LHBs) contains the integral pre-S1 domain, which binds to the HBV receptor sodium taurocholate co transporting polypeptide on the hepatocyte to facilitate viral entry. The accumulation of viral LHBs and its prevalent pre-S mutants in chronic HBV carriers triggers a sustained endoplasmic reticulum (ER) overload response, leading to ER stress-mediated cell proliferation, metabolic switching and genomic instability, which are associated with pro-oncogenic effects. Ground glass hepatocytes identified in HBV-related hepatocellular carcinoma (HCC) patients harbor pre-S deletion variants that largely accumulate in the ER lumen due to mutation-induced protein misfolding and are associated with increased risks of cancer recurrence and metastasis. Moreover, in contrast to the major HBs, which is decreased in tumors to a greater extent than it is in peritumorous regions, LHBs is continuously expressed during tumorigenesis, indicating that LHBs serves as a promising biomarker for HCC in people with CHB. Continuing efforts to delineate the molecular mechanisms by which LHBs regulates pathological changes in CHB patients are important for establishing a correlation between LHBs biomarkers and HCC development.

• endoplasmic reticulum
• ground glass hepatocyte
• hepatitis B virus
• hepatocellular carcinoma
• large surface protein
• pre-S deletion
• sodium taurocholate cotransporting polypeptide
• viral entry

Chronic hepatitis B virus (HBV) infection is a major risk factor for the development of hepatocellular carcinoma (HCC), the leading cause of cancer-related death worldwide. Despite progress in the prevention and therapy of HCC, high incidence and recurrence rates of HCC remain big threats, resulting in poor patient survival. Effective biomarkers and targets of HCC are therefore urgently needed for better management and to improve patient outcomes. Pre-S mutants have been well demonstrated as HBV oncoproteins that play important roles in HCC development through activation of multiple oncogenic signal pathways in hepatocytes, in vitro and in vivo. The presence of pre-S mutants in patients with chronic HBV infection and HBV-related HCC has been associated with a significantly higher risk of HCC development and recurrence after curative surgical resection, respectively. In this review, we summarize the roles of pre-S mutants as biomarkers for predicting HBV-related HCC development and recurrence, and highlight the pre-S mutants-activated oncogenic signal pathways as potential targets for preventing HBV-related HCC development.

• biomarkers
• hepatitis B virus
• hepatocellular carcinoma
• pre-S mutants
• targets

• Antiviral Agents/therapeutic use
• Biomarkers/analysis
• Biomarkers/blood
• Carcinoma, Hepatocellular/diagnosis
• Carcinoma, Hepatocellular/etiology
• Carcinoma, Hepatocellular/prevention & control
• Carcinoma, Hepatocellular/surgery
• Hepatitis B Surface Antigens/analysis
• Hepatitis B Surface Antigens/blood
• Hepatitis B Surface Antigens/genetics
• Hepatitis B virus/genetics
• Hepatitis B, Chronic/complications
• Hepatitis B, Chronic/drug therapy
• Hepatitis B, Chronic/metabolism
• Hepatitis B, Chronic/virology
• Humans
• Liver Cirrhosis/diagnosis
• Liver Cirrhosis/etiology
• Liver Neoplasms/diagnosis
• Liver Neoplasms/etiology
• Liver Neoplasms/prevention & control
• Liver Neoplasms/surgery
• Mutant Proteins/blood
• Mutation
• Neoplasm Recurrence, Local
• Protein Precursors/analysis
• Protein Precursors/blood
• Protein Precursors/genetics
• Risk Factors
• Signal Transduction

• Chronic low-level inflammation
• Cytokines
• HBV
• HCV
• Hepatic sentinel cells
• Hepatocellular carcinoma
• Immune checkpoint blockade
• Immunopathology
• Immunoregulatory mechanisms
• Liver
• Suppressor cells
• T cell exhaustion
• T cells
• Tumour microenvironment
• Wnt/β-catenin

Chronic hepatitis B virus (HBV) infection is a major risk factor for the development of hepatocellular carcinoma (HCC), a leading cause of cancer mortality worldwide. Hepatitis B X protein (HBx) and pre-S2 mutant have been proposed as the two most important HBV oncoproteins that play key roles in HCC pathogenesis. Curcumin is a botanical constituent displaying potent anti-inflammatory and anti-cancer properties without toxic side effects. Phytosomal formulation of curcumin has been shown to exhibit enhanced bioavailability, improved pharmacokinetics, and excellent efficacy against many human diseases. However, effectiveness of phytosomal curcumin for HCC treatment remains to be clarified. In this study, we evaluated chemopreventive effect of phytosomal curcumin on HBV-related HCC by using a transgenic mouse model specifically expressing both HBx and pre-S2 mutant in liver. Compared with unformulated curcumin, phytosomal curcumin exhibited significantly greater effects on suppression of HCC formation, improvement of liver histopathology, decrease of lipid accumulation and leukocyte infiltration, and reduction of total tumor volume in transgenic mice. Moreover, phytosomal curcumin exerted considerably stronger effects on activation of anti-inflammatory PPARγ as well as inhibition of pro-inflammatory NF-κB than unformulated curcumin. Furthermore, phytosomal curcumin showed a comparable effect on suppression of oncogenic mTOR activation to unformulated curcumin. Our data demonstrated that phytosomal curcumin has promise for HCC chemoprevention in patients with chronic HBV infection.

Background: β2-adrenoceptors (β2-ARs) are expressed on the surface of immune cells, including tumor-associated macrophages (TAMs). Previous studies have demonstrated that the expression of β2-ARs in hepatocellular carcinoma (HCC) is significantly increased in vitro. However, the role of β2-AR in M2-polarized macrophages remains unclear. G protein-coupled receptor kinase 2 (GRK2) can regulate G protein-coupled receptor (GPCR). Previous studies showed that down-regulation of GRK2 in HCC contributes the HCC progression, but it still remains unclear whether the regulation of β2-AR in M2-polarized macrophages by GRK2 can promote HCC.

Purpose: The present study was designed to investigate the role of activated β2-AR in M2-polarized macrophages in the HCC progression and GRK2 regulatory effect, as well as the underlying mechanisms involved.

Results: The results demonstrated that the M2-polarized macrophages were increased with HCC progression. In vitro, the activation of β2-AR by terbutaline in M2-polarized macrophages elevated the proliferative, migratory and invasive attributes of HCC cells. Furthermore, GRK2 down-regulation in β2-AR activated M2-polarized macrophages activated the downstream cyclic adenosine monophosphate (cAMP)/protein kinase A/cAMP-response element binding protein and cAMP/interleukin-6/signal transducer and the activator of transcription 3 signaling pathways, contributing to the secretion of tumor-associated cytokines, and thus resulting in the promotion of malignant biological behavior in HCC cells.

Conclusion: These findings suggest that the regulation of β2-AR occurs through the silencing of GRK2 in M2-polarized macrophages, which is conducive to HCC development, through its engagement in the activation of downstream signaling.

• G protein-coupled receptor kinase 2
• beta2-adrenoceptor
• hepatocellular carcinoma
• macrophage
• therapeutic target
• tumor microenvironment

Silybin is a flavonolignan extracted from Silybum marianum (milk thistle) with hepatoprotective, antioxidant, and anti-inflammatory activity. Several studies have shown that silybin is highly effective to prevent and treat different types of cancer and that its antitumor mechanisms involve the arrest of the cell cycle and/or apoptosis. An MTT assay was performed to study cell viability, lipid peroxidation, extracellular NO production, and scavenger enzyme activity were studied by Thiobarbituric Acid-Reactive Species (TBARS) assay, NO assay, and MnSOD assay, respectively. Cell cycle and apoptosis analysis were performed by FACS. miRNA profiling were evaluated by real time PCR. In this study, we demonstrated that Silybin induced growth inhibition blocking the Hepg2 cells in G1 phase of cell cycle and activating the process of programmed cell death. Moreover, the antiproliferative effects of silybin were paralleled by a strong increase of the number of ceramides involved in the modulation of miRNA secretion. In particular, after treatment with silybin, miR223-3p and miR16-5p were upregulated, while miR-92-3p was downregulated (p < 0.05). In conclusion, our results suggest that silybin-Induced apoptosis occurs in parallel to the increase of ceramides synthesis and miRNAs secretion in HepG2 cells.

• PTEN
• apoptosis
• ceramides
• hepatocarcinoma
• miRNAs
• silybin
• sorafenib

• Antineoplastic Agents, Phytogenic/pharmacology
• Apoptosis/drug effects
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/metabolism
• Cell Cycle Checkpoints/drug effects
• Cell Line, Tumor
• Cell Survival/drug effects
• Ceramides/biosynthesis
• Dose-Response Relationship, Drug
• Gene Expression Profiling
• Gene Expression Regulation, Neoplastic/drug effects
• Humans
• Lipid Metabolism/drug effects
• Liver Neoplasms/genetics
• Liver Neoplasms/metabolism
• MicroRNAs/genetics
• Nitric Oxide/biosynthesis
• Silybin/pharmacology

Ground glass hepatocytes (GGHs), a histological hallmark of chronic hepatitis B virus (HBV) infection, contain excessive hepatitis surface antigen (HBsAg) in the endoplasmic reticulum (ER), which is linked to unfolded protein response (UPR). The mechanism by which HBV activates UPR has not been fully defined. To investigate this, HepG2-NTCP cells and primary human hepatocytes (PHHs) were either infected with HBV or transduced with adenoviral vectors expressing replication-competent HBV genome or individual HBV genes. UPR markers were evaluated by qPCR, Western blotting, and immunofluorescence. Apoptosis and cell viability were measured by Caspase-3/7 and ATPlite assay respectively. We found that UPR markers were induced by the overexpression of HBsAg in HepG2-NTCP cells and PHHs. Elevation of UPR-induced genes showed a dose-dependent correlation with HBsAg levels. In HBV-infected livers, GGHs also demonstrated excessive accumulation of HBsAg associated with increased BIP/GRP78 staining, a marker of UPR. Prolonged activation of UPR by HBsAg overexpression induced signs of apoptosis. Overexpression of HBsAg can induce ER stress through protein kinase RNA-like endoplasmic reticulum kinase (PERK) pathway in vitro, and may be linked to the appearance of GGHs. The activation of UPR by HBsAg may sensitize hepatocytes to cell death and result in possible subsequent cellular changes leading to a premalignant phenotype.

• apoptosis
• chronic hepatitis B
• endoplasmic reticulum stress
• ground glass hepatocyte
• hepatocellular carcinoma
• liver disease

• Adenoviridae/genetics
• Apoptosis
• Carcinoma, Hepatocellular/pathology
• Carcinoma, Hepatocellular/virology
• Cell Survival
• Cells, Cultured
• Endoplasmic Reticulum/virology
• Endoplasmic Reticulum Chaperone BiP
• Hep G2 Cells
• Hepatitis B Surface Antigens/genetics
• Hepatitis B virus/chemistry
• Hepatitis B virus/genetics
• Hepatitis B, Chronic/pathology
• Hepatocytes/pathology
• Hepatocytes/virology
• Humans
• Liver Neoplasms/pathology
• Liver Neoplasms/virology
• Unfolded Protein Response

• Cellular function
• Hepatocellular carcinoma
• Non-coding RNAs
• The mTOR pathway
• Therapeutic target

• Animals
• Carcinoma, Hepatocellular/metabolism
• Humans
• Liver Neoplasms/metabolism
• Signal Transduction/physiology
• TOR Serine-Threonine Kinases/metabolism

• Hepatitis B virus
• Hepatocellular carcinoma (HCC)
• Vascular endothelial growth factor (VEGF)
• Vascular endothelial growth factor receptor 2 (VEGFR2)
• Woodchuck

• Animals
• COS Cells
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/metabolism
• Carcinoma, Hepatocellular/pathology
• Chlorocebus aethiops
• Humans
• Liver Neoplasms, Experimental/genetics
• Liver Neoplasms, Experimental/metabolism
• Liver Neoplasms, Experimental/pathology
• Marmota/genetics
• Marmota/metabolism
• Neoplasm Proteins/genetics
• Neoplasm Proteins/metabolism
• Neovascularization, Pathologic/genetics
• Neovascularization, Pathologic/metabolism
• Neovascularization, Pathologic/pathology
• Vascular Endothelial Growth Factor A/genetics
• Vascular Endothelial Growth Factor A/metabolism

• Acute-phase proteins
• Acute-phase response
• Alpha1-antitrypsin deficiency
• Autophagy
• ER folding machinery
• ER stress response
• ER-associated degradation
• Fibrinogen storage disease
• Hepatitis B infection
• Proteotoxic stress
• Secretory proteins

Several viruses are recognized as the direct or indirect causative agents of human tumors and other severe human diseases. Vascular endothelial growth factor (VEGF) is identified as a principal proangiogenic factor that enhances the production of new blood vessels from existing vascular network. Therefore, oncogenic viruses such as Kaposi’s sarcoma herpesvirus (KSHV) and Epstein-Barr virus (EBV) and non-oncogenic viruses such as herpes simplex virus (HSV-1) and dengue virus, which lack their own angiogenic factors, rely on the recruitment of cellular genes for angiogenesis in tumor progression or disease pathogenesis. This review summarizes how human viruses exploit the cellular signaling machinery to upregulate the expression of VEGF and benefit from its physiological functions for their own pathogenesis. Understanding the interplay between viruses and VEGF upregulation will pave the way to design targeted and effective therapeutic approaches for viral oncogenesis and severe diseases.

• Dengue fever virus
• Epstein-Barr virus (EBV)
• Hantavirus
• Hepatitis B Viruses (HBV)
• Hepatitis C Viruses (HCV)
• Herpes Simplex Virus (HSV)
• Kaposi’s sarcoma herpesvirus (KSHV)
• angiogenesis
• therapeutics
• vascular endothelial growth factor (VEGF)
• vascular endothelial growth factor receptor (VEGFR)

Chronic hepatitis B is a global health problem. The clinical outcomes of chronic hepatitis B infection include asymptomatic carrier state, chronic hepatitis (CH), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). Because of the spontaneous error rate inherent to viral reverse transcriptase, the hepatitis B virus (HBV) genome evolves during the course of infection under the antiviral pressure of host immunity. The clinical significance of pre-S/S variants has become increasingly recognized in patients with chronic HBV infection. Pre-S/S variants are often identified in hepatitis B carriers with CH, LC, and HCC, which suggests that these naturally occurring pre-S/S variants may contribute to the development of progressive liver damage and hepatocarcinogenesis. This paper reviews the function of the pre-S/S region along with recent findings related to the role of pre-S/S variants in liver diseases. According to the mutation type, five pre-S/S variants have been identified: pre-S deletion, pre-S point mutation, pre-S1 splice variant, C-terminus S point mutation, and pre-S/S nonsense mutation. Their associations with HBV genotype and the possible pathogenesis of pre-S/S variants are discussed. Different pre-S/S variants cause liver diseases through different mechanisms. Most cause the intracellular retention of HBV envelope proteins and induction of endoplasmic reticulum stress, which results in liver diseases. Pre-S/S variants should be routinely determined in HBV carriers to help identify individuals who may be at a high risk of less favorable liver disease progression. Additional investigations are required to explore the molecular mechanisms of the pre-S/S variants involved in the pathogenesis of each stage of liver disease.

• Hepatitis B virus
• Pre-S/S mutant
• Pre-S deletion
• Splice variant
• spPS1
• Chronic hepatitis
• Liver cirrhosis
• Hepatocellular carcinoma

• Carcinoma, Hepatocellular/pathology
• Carcinoma, Hepatocellular/virology
• Carrier State/pathology
• Carrier State/virology
• DNA, Viral/genetics
• Disease Progression
• Endoplasmic Reticulum Stress
• Genotype
• Hepatitis B Surface Antigens/genetics
• Hepatitis B virus/genetics
• Hepatitis B virus/pathogenicity
• Hepatitis B, Chronic/pathology
• Hepatitis B, Chronic/virology
• Humans
• Liver/pathology
• Liver/virology
• Liver Cirrhosis/pathology
• Liver Cirrhosis/virology
• Liver Neoplasms/pathology
• Liver Neoplasms/virology
• Mutation
• Protein Precursors/genetics
• RNA Splicing/genetics
• RNA, Viral/genetics
• Viral Envelope Proteins/genetics
• Virus Replication/genetics

Hepatitis B virus (HBV) is a driver of hepatocellular carcinoma, and two viral products, X and large surface antigen (LHBS), are viral oncoproteins. During chronic viral infection, immune-escape mutants on the preS2 region of LHBS (preS2-LHBS) are gain-of-function mutations that are linked to preneoplastic ground glass hepatocytes (GGHs) and early disease onset of hepatocellular carcinoma. Here, we show that preS2-LHBS provoked calcium release from the endoplasmic reticulum (ER) and triggered stored-operated calcium entry (SOCE). The activation of SOCE increased ER and plasma membrane (PM) connections, which was linked by ER- resident stromal interaction molecule-1 (STIM1) protein and PM-resident calcium release- activated calcium modulator 1 (Orai1). Persistent activation of SOCE induced centrosome overduplication, aberrant multipolar division, chromosome aneuploidy, anchorage-independent growth, and xenograft tumorigenesis in hepatocytes expressing preS2- LHBS. Chemical inhibitions of SOCE machinery and silencing of STIM1 significantly reduced centrosome numbers, multipolar division, and xenograft tumorigenesis induced by preS2-LHBS. These results provide the first mechanistic link between calcium homeostasis and chromosome instability in hepatocytes carrying preS2-LHBS. Therefore, persistent activation of SOCE represents a novel pathological mechanism in HBV-mediated hepatocarcinogenesis.

• ER stress
• SOCE
• aneuploidy
• ground-glass hepatocytes
• hepatitis B virus

• anti-viral therapy
• ground glass hepatocytes
• hepatitis B virus
• hepatocellular carcinoma
• pre-S mutation

• U-shaped distribution
• deletion index
• hepatocarcinogenesis
• meta-analysis
• next-generation sequencing

• Chronic hepatitis
• Genotype
• Hepatitis B virus
• Hepatocellular carcinoma
• Liver cirrhosis
• Pre-S deletion

• hepatitis b virus (hbv)
• hepatocellular carcinoma (hcc)
• ground glass hepatocytes (gghs)
• pre-s2 mutant
• mammalian target of rapamycin (mtor)

• Animals
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/metabolism
• Carcinoma, Hepatocellular/virology
• Hepatitis B virus/pathogenicity
• Hepatocytes/metabolism
• Humans
• Liver Neoplasms/genetics
• Liver Neoplasms/metabolism
• Liver Neoplasms/virology
• Signal Transduction/genetics
• Signal Transduction/physiology
• TOR Serine-Threonine Kinases/genetics
• TOR Serine-Threonine Kinases/metabolism