A score model for predicting post-liver transplantation survival in HBV cirrhosis-related hepatocellular carcinoma recipients: a single center 5-year experience

BACKGROUND： The prognostic prediction of liver transplantation（LT） guides the donor organ allocation. However, there is currently no satisfactory model to predict the recipients’ outcome, especially for the patients with HBV cirrhosis-related hepatocellular carcinoma（HCC）. The present study was to develop a quantitative assessment model for predicting the post-LT survival in HBV-related HCC patients.METHODS： Two hundred and thirty-eight LT recipients at the Liver Transplant Center, First Affiliated Hospital, Zhejiang University School of Medicine between 2008 and 2013 were included in this study. Their post-LT prognosis was recorded and multiple risk factors were analyzed using univariate and multivariate analyses in Cox regression.RESULTS： The score model was as follows： 0.114×（Child-Pugh score）-0.002×（positive HBV DNA detection time）＋0.647×（number of tumor nodules）＋0.055×（max diameter of tumor nodules）＋0.231×ln AFP＋0.437×（tumor differentiation grade）.The receiver operating characteristic curve analysis showed that the area under the curve of the scoring model for predicting the post-LT survival was 0.887. The cut-off value was 1.27, which was associated with a sensitivity of 72.5% and a specificity of 90.7%, respectively.CONCLUSION： The quantitative score model for predicting post-LT survival proved to be sensitive and specific.

Quantitative analysis of plasma HBV DNA for early evaluation of the response to transcatheter arterial embolization for HBV-related hepatocellular carcinoma

AIM： To assesse changes in plasma HBV DNA after TAE in HBV-related HCC and correlate the levels with the pattern of lipiodol accumulation on CT. METHODS： Between April and June 2001, 14 patients with HBV-associated HCC who underwent TAE for inoperable or recurrent tumor were studied. Levels of plasma HBV DNA were measured by real-time quantitative PCR daily for five consecutive days after TAE. More than twofold elevation of circulating HBV DNA was considered as a definite elevation. Abdominal CT was performed 1-2 mo after TAE for the measurement of lipiodol retention. RESULTS： Circulating HBV DNA in 10 out of 13 patients was elevated after TAE, except for one patient whose plasma HBV DNA was undetectable before and after TAE. In group Ⅰ patients （n = 6）, the HBV DNA elevation persisted for more than 2 d, while in group Ⅱ （n = 7）, the HBV DNA elevation only appeared for i d or did not reach a definite elevation. There were no significant differences in age or tumor size between the two groups. Patients in group Ⅰ had significantly better lipiodol retention （79.31±28.79%） on subsequent abdominal CT than group Ⅱ （18.43±10.61%） （P = 0.02）. CONCLUSION： Patients with durable HBV DNA elevation for more than 2 d correlated with good lipiodol retention measured 1 mo later, while others associated with poor lipiodol retention. Thus, circulating HBV DNA may be an early indicator of the success or failure of TAE.

The significance of SREBP1 expression in HBV-related hepatocellular carcinoma

Objective:To investigate the relationship between SREBP1 protein expression and clinicopathological features of hepatitis B virus infection-related hepatocellular carcinoma.Methods:A total of 91 cases of hepatocellular carcinoma associated with hepatitis B virus infection and paired adjacent liver tissue samples were collected from the Department of Pathology,the First Affiliated Hospital of Hainan Medical College.The expression of SREBP1 protein in cancer and paired adjacent tissues was detected by immunohistochemical staining.The relationship between SREBP1 protein expression and clinicopathological features of hepatocellular carcinoma was analyzed by chi-square test.Results:The results of immunohistochemical staining showed that the ex-pression of SREBP1 protein in hepatocellular carcinoma was significantly higher than that in adjacent liver tissues(P<0.001).The expression level of SREBP1 protein was significantly correlated with tumor differentiation,distant metastasis or local recurrence in patients with hepatocellular carcinoma(P<0.05).The expression of SREBP1 protein was significantly up-regulated in the high HBV-related HCC virus load group(>103 IU/mL)compared with the low HBV-related HCC virus load group 3(0~10 IU/mL)(P<0.05).Conclusion:1.The expression of SREBP1 protein is correlated with hepatocellular carcinoma,and shows a high expression trend,which provides direction and theoretical basis for the study of lipid metabolism regulation mechnism related to the occurrence and development of hepatocellular carcinoma.2.The expression of SREBP1 protein is correlated with the replication activity of hepatitis B virus,which provides a new research direction for the exploration of the mechanism of the occurrence and development of hepatitis B virus infection-related liver cancer.

Abnormal expression of HSP gp96 associated with HBV replication in human hepatocellular carcinoma

BACKGROUND: Heat shock protein (HSP) gp96 is a member of the HSP90 family and presumably overexpresses as a result of stimulation by mutated or abnormal proteins. Its abnormal expression correlates with carcinogenesis, progression and prognosis of hepatocellular carcinoma (HCC). In this study, we investigated the pathological characteristics of liver gp96 expression and its relationship with hepatitis B virus (HBV) replication in HCC patients. METHODS: Tumor specimens were prospectively collected from 30 HCC patients undergoing liver resection. Total RNAs were extracted from HCC or their noncancerous tissues. The distribution of gp96 expression in hepatocytes was investigated by streptavidin peroxidase (S-P) immunohistochemistry and tissue HBV-DNA was detected by the in situ molecular hybridization technique. The association of gp96 expression with HBV replication, and the histopathological characteristics of HCC were analyzed. RESULTS: The gp96 was strongly expressed in HCC (73.3%, 22 of 30) and weakly (46.7%, 14 of 30) in non-cancerous tissues. The gp96 expression in HCC tissues was correlated with degree of tumor differentiation and tumor size, but not with tumor number (P>0.05). Immunohistochemical analysis showed that 17 of 19 HCC patients with HBVDNA -positive were strongly expressed for gp96, whereas only 5 of 11 patients with HBV-DNA-negative were positive for gp96. A significant difference was found between the two groups (89.5% vs. 45.5%, P<0.05). CONCLUSIONS: The abnormal expressions of HSP gp96 in HCC tissues are associated with HBV replication. This finding indicates that HBV infection plays an important role in the development of HCC.

MCM3AP,a Novel HBV Integration Site in Hepatocellular Carcinoma and Its Implication in Hepatocarcinogenesis

A novel HBV integration site involved in hepatocarcinogenesis was investigated. The HBV DNA integration sites were detected by Alu-PCR in hepatocellular carcinoma tissues, matched surrounding liver tissues in 30 patients with hepatitis B-related hepatocellular carcinoma (HCC) and 3 cases of normal liver tissues. The integration sites and flanking sequences in human genome were sequenced and blasted, and the expression of integrated HBV genes was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The influence of the up-regulated expression of integrated genes on hepatocarcinogenesis was analyzed. Nineteen integration sites of HBV DNA into HCC tissues were obtained by RT-PCR and sequencing. These genes encoding proteins were: LOC51030, LOC157777, minichromosome maintenance complex component 3 associated protein (MCM3AP), MCTP1, SH3 and multiple ankyrin repeat domains 2 isoform 2, CCDC40, similar to HCG2033532, mitochondrial ribosomal S5 pseudogene 4. One of them was integrated into the intron of MCM3AP. RT-PCR demonstrated that the expression levels of MCM3AP mRNA in HCC tissues, matched surrounding liver tissues and normal liver tissues were in a descendent order. The ratio of MCM3AP mRNA to the GAPDH mRNA in these three tissues was 1.07375, 0.21573, 0.06747 respectively, with the difference being statistically significant among them (P<0.05). Meanwhile, the expression levels of MCM3AP mRNA from HCC tissues in which HBV DNA integrated into MCM3AP were still significantly higher than those without HBV DNA integrated into MCM3AP. It was concluded that the HBV DNA integration sites into human genome were random, and MCM3AP was a new site. The up-regulated MCM3AP mRNA may affect flanking sequences which promote the hepatocarcinogenesis.

Synergistic Action of Clonorchiasis,HBV Infection and Alcohol Consumption on Primary Hepatocellular Carcinoma

OBJECTIVE It has been recognized that HBV infection and alcohol consumption are two important risk factors for primary hepatocellular carcinoma （HCC）. Recently, the role of clonorchiasis as a risk factor for HCC is controversial. We aimed to investigate whether these factors increase the risk of HCC in Guangxi, China. METHODS A hospital-based, case-control study of HCC was conducted from July 2005 to July 2007. We enrolled 500 consecutive patients with HCC as an experimental group and 500 patients without tumor in liver as a control group. The risk factors that the patients were exposed to were assessed. RESULTS Comparing the risks of developing the HCC, we found out the following results. The risk of developing HCC for the patients with clonorchiasis was 5 folds of that for the patients without clonorchiasis （OR = 5.0; 95% CI： 3.1-8.1）, and the risk for the patients with alcohol consumption was 3 folds of that for the patients without drinking alcohol （OR = 3.4; 95% CI： 2.3-4.9）, and similarly, the risk for the patients with HBV infection was 21 times of that for the patients without HBV infection （OR = 20.6; 95% CI： 14.3-29.7）. According to crossover analysis, there was significant interaction among clonorchiasis, HBV infection and alcohol consumption, with synergistic indices greater than 1. The etiologic fractions attributed to these interactions [EF （A × B）] are 0.7465, 0.5789 and 0.5506, respectively. CONCLUSION Clonorchiasis, HBV infection and heavy alcohol consumption are independent risk factors for developing HCC in our population in Guangxi, and as they can interact synergistically, the risk of developing HCC is increased. Data from this study may indicate new prevention strategies of developing HCC in high-risk individuals.

THE EXPRESSIONS OF HBV X GENE AND ets-2, IGF-Ⅰ, c-myc AND N-ras ONCOGENES IN HUMAN HEPATOCELLULAR CARCINOMA AND TUMOR-ADJACENT TISSUES

The expressions of HBV X gene and ets-2, IGF-I, c-myc and N-ras were studied in 7 pairs of human primary hepatocellular carcinoma (PHC) and tumor-adjacent tissues, using RNA hybridization and im-munoblot methods. The results showed that specific 17 and 28 kD HBV X gene products (HBxAg) were existed in a portion of PHC and tumor-adjacent tissues. The 17 kD HBxAg was detected in the sera of 3 patients who also had 17 kD HBxAg in their liver tissues. Multiple expressions of oncogenes such as ets-2, c-myc and N-ras were observed in PHC and tumor-adjacent tissues that had HBxAg expressed, indicating HBxAg might function as a transactivator in the course of intracellular proto-oncogene activation. It is also observed that in some tumor-adjacnet tissues the expressions of ets-2, c-myc and N-ras were higher than those in corresponding PHC. The relationship of HBxAg to the expression of est-2, IGF-Ⅱ, c-myc and their possible roles in the carcinogenesis of PHC are discussed.

The function of HBx in HBV-induced hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is the second cause cancer death in the world. HCC is frequently diagnosed at advanced stages with intrahepatic metstasis or vascular invasion and has a poor prognosis with a high mortality rate. In the world, hepatitis B virus(HBV) caused over 50% HCC, making it the most common carcinogen after tobacco, and the encoded protein of the HBV X gene(HBx) plays a critical role in the tumorigenesis of HBV-related HCC. There is a lot of literature about the function of HBx. In this article we summary the main function of HBx in HCC.

HBV Infection Promotes the Occurrence and Development of Hepatocellular Carcinoma through Impairing the Inhibitory Effect of PPP2R5A on MAPK/AKT/WNT Signaling Pathway

Reversible phosphorylation and dephosphorylation play important roles in cell function and cell signal transduction. PPP2R5A (protein phosphatase 2 regulatory subunit B’ alpha) is responsible for specifically regulating the catalytic function, substrate specificity and intracellular localization of the tumor suppressor phosphatase PP2A (serine/threonine protein phosphatase 2A). Therefore, the abnormal expression and function of PPP2R5A may be related to canceration. The aim of this study was to reveal its role in the occurrence and development of hepatocellular carcinoma (HCC). It is hoped that the results of this study can provide guidance for the prevention and treatment of HCC. The results showed that PPP2R5A inhibited the proliferation and metastasis of HCC cells, and acted as a tumor suppressor in HCC cells, but it had no significant effect on cell cycle. Further research found that PPP2R5A exerted tumor suppressor efficacy by inhibiting the MAPK/AKT/WNT signaling pathway. Combined with analysis of clinical tissue samples and TCGA database, it was found that the expression of PPP2R5A in tumor tissues of Chinese HCC patients was down-regulated and significantly correlated with the progression-free survival (PFS) of HCC patients. On the contrary, PPP2R5A showed an up-regulation trend in HCC cases in TCGA database although its effect on PFS was the same with that in Chinese HCC patients. Hepatitis B virus (HBV) infection is the main pathogenic factor of HCC in China. It was found that HBV infection reduced the content of PPP2R5A in cells. It was concluded that HBV inhibited the initiation of the protective mechanism mediated by PPP2R5A, making the occurrence and progress of HCC more “unimpeded”. This conclusion will further reveal the role of PPP2R5A in HBV-induced and HBV-unrelated HCC, therefore, providing clues for the prevention and treatment of the two types of HCC, respectively.

抗病毒治疗在预防HBV DNA阴性的肝细胞癌术后复发中的影响

目的探讨抗病毒药物治疗对乙肝病毒(HBV)DNA阴性的肝细胞癌(HCC)患者预防术后复发的影响。方法80例HBV DNA阴性的HCC患者随机分为两组。抗病毒组给予手术切除+术后常规预防性经肝动脉化疗栓塞术(TACE)+术前预防性抗病毒治疗,40例;对照组给予手术切除+术后常规预防性TACE治疗,40例。术后随访2年,比较两组患者术后HBV再激活率,观察两组患者术后6个月、1年、2年复发率。结果术后随访2年,对照组出现HBV再激活29例(72.50%),抗病毒组出现HBV再激活3例(7.50%),两组HBV再激活率比较,差异有统计学意义(χ^(2)=5.43,P<0.05)。抗病毒组术后6个月、1年、2年复发率均明显低于对照组,差异有统计学意义(χ^(2)分别=8.35、11.61,P均<0.05)。多因素分析结果显示:给予抗病毒治疗是影响HCC术后复发的保护因素(OR=0.27,P<0.05)。结论对于HBV DNA阴性的HCC患者进行预防性抗病毒治疗,能有效减少HBV再激活,有助于降低术后2年内的肿瘤复发率。

Expression of nuclear factor-KB in hepatocellular carcinoma and its relation with the X protein of hepatitis B virus

AIM In this study we investigated the relationship of the X protein of HBV and nuclear factor-κB(NF-κB)and the expression of NF-κB in human hepatocellular carcinoma tissues. METHODS Immunohistochemistry SP method was used to detect the expression of NF-κB and the X protein of HBV in human hepatocellular carcinoma tissues of 52 cases.Gene transfection mediated by lipofectamine was used to transfect the eukaryotic expression vector pCDNA3.1-HBX of HBV x gene into human hepatocellular carcinoma cell line HCC-9204 and NF-κB was detected. RESULTS NF-κB was widely expressed in human hepatocellular carcinoma tissues in a total of 52 cases and its expression was related to the X protein of HBV.NF-κB was localized both in the cytoplasm and the nuclei of hepatocellular carcinoma cells in 11 cases which were positive for the X protein of HBV while in 41 cases negative for the X protein of HBV,NF-κB was only localized in the cytoplasm of hepatocellular carcinoma cells but translocated to the nuclei of hepatocellular carcinoma cells after the eukaryotic expression vector pCDNA3.1-HBX was transfected into HCC-9204 cells. CONCLUSION This study strongly suggests that the nuclear factor NF-κB is widely expressed in hepatocellular carcinoma tissues in different styles according to the expression of the X protein of HBV.NF-κB is abnormally activated in hepatocellular carcinoma,which is probably rélated to the X protein of HBV.The X protein of HBV can activate NF-κB to translocate into nuclei of hepatocellular carcinoma cells.

Quantitative evaluation of hepatitis B virus mutations and hepatocellular carcinoma risk:a meta-analysis of prospective studies

Background： The temporal relationship between hepatitis B virus （HBV） mutations and hepatocellular carcinoma （HCC） remains unclear. Methods： We conducted a meta-analysis including cohort and nested case-control studies to prospectively examine the HCC risk associated with common variants of HBV in the PreS, Enhancer Ⅱ, basal core promoter （BCP） and precore regions. Pertinent studies were identified by searching PubMed, Web of Science and the Chinese Biological Medicine databases through to November 2014. Study-specific risk estimates were combined using fixed or random effects models depending on whether significant heterogeneity was detected. Results： Twenty prospective studies were identified, which included 8 cohort and 12 nested case-control studies. There was an increased risk of HCC associated with any PreS mutations with a pooled relative risk （RR） of 3.82 [95% confidence interval （CI）： 2.59-5.61]. The pooled-RR for PreS deletion was 3.98 （95% CI： 2.28-6.95）, which was higher than that of PreS2 start codon mutation （pooled-RR=2.63, 95% CI： 1.30-5.34）. C1653T in Enhancer Ⅱ was significantly associated with HCC risk （pooled-RR=l.83; 95% CI： 1.21-2.76）. For mutations in BCP, statistically significant pooled-RRs of HCC were obtained for T1753V （pooled- RR=2.09; 95% CI： 1.49-2.94） and AI762T/G1764A double mutations （pooled-RR=3.11; 95% CI： 2.08- 4.64）. No statistically significant association with HCC risk was observed for G1896A in the precore region （pooled-RR=0.77; 95% CI： 0.47-1.26）. Conclusions： This study demonstrated that PreS mutations, C1653T, T1753V, and A1762T/G1764A, were associated with an increased risk of HCC. Clinical practices concerning the HCC risk prediction and diagnosis may wish to focus on patients with these mutations.

Hepatitis B virus genotypes and hepatocellular carcinoma in Thailand

AIM: The role of hepatitis B virus (HBV) genotypes on the clinical features and prognosis of patients with hepatocellular carcinoma (HCC) is currently unknown. The aim of the present study was to evaluate the distribution of HBV genotypes and their clinical relevance in Thai patients.METHODS: HBV genotypes were determined by PCR-RFLP in stored sera of 93 asymptomatic carriers, 103 patients with chronic hepatitis, 60 patients with cirrhosis and 76patients with HCC. The clinical data were analyzed in relation to the HBV genotype.RESULTS: HBV genotypes C and B were predominant in Thailand, accounting for 73% and 21%, respectively. The distributions of genotypes B and C were similar in HCC patients compared to the other groups. Genotype C was significantly more common in HCC patients who were under 40 years old than genotype B (18% vs 0%, P= 0.03), but was significantly less common in patients older than 60 years (26% vs 56.5%, P= 0.01). The positive rate of hepatitis B e antigen (HBeAg) in patients with genotype C was significantly higher than that in patients with genotype B (71.6% vs 44.4%, P = 0.03 in chronic hepatitis; 56.8% vs 11.1%,P = 0.01 in cirrhosis). There were no differences between HCC patients with genotypes B and C regarding tumor staging by CLIP criteria and the overall median survival. Multivariate analyses showed that HBV genotype was not an independent prognostic factor of survival in HCC patients.CONCLUSION: Patients with genotype C had a higher positive rate of HBeAg and exhibited earlier progression of cirrhosis and HCC than those with genotype B. However,there were no differences in the risk of developing HCC and its prognosis between patients with these genotypes.

Hepatocellular carcinoma in Egypt: A single center study over a decade

AIM： To identify the trend, possible risk factors and any pattern change of hepatocellular carcinoma （HCC） in Egypt over a decade. METHODS： All HCC patients attending Cairo Liver Center between January 1993 and December 2002, were enrolled in the study. Diagnosis of HCC was based on histopathological examination and/or detection of hepatic focal lesions by two imaging techniques plus α-fetoprotein level above 200 ng/mL. The duration of the study was divided into two periods of 5 years each; period Ⅰ （1993-1997） and period Ⅱ （1998-2002）. Trend, demographic features of patients （age, gender, and residence）, risk factors （HBsAg, HCV-Ab, schistosomiasis and others） and pattern of the focal lesions were compared between the two periods. Logistic regression model was fitted to calculate the adjusted odds ratios for the potential risk factors. The population attributable risk percentage was calculated to estimate the proportion of HCC attributed to hepatitis B and C viral infections. RESULTS： Over a decade, 1 328 HCC patients out of 22 450 chronic liver disease （CLD） patients were diagnosed with an overall proportion of 5.9%. The annual proportion of HCC showed a significant rising trend from 4.0% in 1993 to 7.2% in 2002 （P = 0.000）. A significant increase in male proportion from 82.5% to 87.6% （P = 0.009）; M/F from 5：1 to 7：1 and a slight increase of the predominant age group （40-59 years） from 62.6% to 66.8% （P = 0.387）in periods Ⅰ and Ⅱ respectively, reflecting a shift to younger age group. In the bivariate analysis, HCC was significantly higher in rural residents, patients with history of schistosomiasis and/or blood transfusion. Yet, after adjustment, these variables did not have a significant risk for development of HCC. There was a significant decline of HBsAg from 38.6% to 20.5% （P = 0.000）, and a slight increase of HCV-Ab from 85.6% to 87.9% in periods I and II respectively. HBV conferred a higher risk to develop HCC more than HCV in period Ⅰ （OR 1.9 vsl.6） and period Ⅱ （OR 2.7 vs 2.0）, but the relative contribution of HBV for development of HCC declined in period Ⅱ compared to period Ⅰ （PAR% 4.2%, 21.32%）. At presentation, diagnostic α-fetoprotein level （≥200 ng/mL） was demonstrated in 15.6% vs28.9% and small HCC （≤3 cm） represented 14,9% vs 22,7% （P = 0,0002） in periods Ⅰ and Ⅱ respectively. CONCLUSION： Over a decade, there was neady a twofold increase of the proportion of HCC among CLD patients in Egypt with a significant decline of HBV and slight increase of HCV as risk factors. α-Fetoprotein played a limited role in diagnosis of HCC, compared to imaging techniques. Increased detection of small lesions at presentation reflects increased awareness of the condition.

Liver expression of Nrf2-related genes in different liver diseases

BACKGROUND： The KEAP1-Nrf2 antioxidant signaling pathway is important in protecting liver from various insults. However,little is known about the expression of Nrf2-related genes in human liver in different diseases.METHODS： This study utilized normal donor liver tissues（n=35）, samples from patients with hepatocellular carcinoma（HCC, n=24）, HBV-related cirrhosis（n=27）, alcoholic cirrhosis（n=5） and end-stage liver disease（n=13）. All of the liver tissues were from the Oriental Liver Transplant Center, Beijing,China. The expressions of Nrf2 and Nrf2-related genes, including its negative regulator Kelch-like ECH-associated protein 1（KEAP1）, its targeted gene NAD（P）H-quinone oxidoreductase 1（NQO1）, glutamate-cysteine ligase catalytic subunit（GCLC） and modified subunit（GCLM）, heme oxygenase 1（HO-1） and peroxiredoxin-1（PRDX1） were evaluated. RESULTS： The expression of Nrf2 was decreased in HCC, increased in alcoholic cirrhosis and end-stage liver disease. The expression of KEAP1 was increased in all of the liver samples.The most notable finding was the increased expression of NQO1 in HCC（18-fold）, alcoholic cirrhosis（6-fold）, endstage liver disease（5-fold） and HBV-related cirrhosis（3-fold）.Peri-HCC also had 4-fold higher NQO1 m RNA as compared to the normal livers. GCLC m RNA levels were lower only in HCC, as compared to the normal livers and peri-HCC tissues.GCLM m RNA levels were higher in HBV-related cirrhosis and end-stage liver disease. HO-1 m RNA levels were increased in all liver tissues except for HCC. Peri-HCC had higher PRDX1 m RNA levels compared with HCC and normal livers.CONCLUSION： Nrf2 and Nrf2-related genes are aberrantly expressed in the liver in different diseases and the increase of NQO1 was the most notable finding, especially in HCC.

Biochemical overview of the recent findings on the correlation between viral hepatitis and its related hepatocellular carcinoma

As one of the most common primary liver cancers, hepatocellular carcinoma(HCC) usually occurs in the presence of inflammation. Compared to other risk factors such as alcohol abuse, aflatoxin, and obesity, virus-induced hepatitis can be effectively prevented by vaccines. For the past several decades, HCC has been believed to be closely related to viral infections although no comprehensive mechanism was established regarding the contribution of viral hepatitis toward HCC. Recent studies have shown that viral infection plays multiple roles in the process of carcinogenesis by causing an increase in genomic instability, cancer-promoting genetic mutations, signal pathway interruption, and tumor suppressor gene inhibition. Sorafenib has become a novel option for HCC patients, especially those who are in advanced disease stage for which conventional treatment methods are not recommended. Future studies should focus more on novel targeted drugs which can be adopted as alternatives to sorafenib or as second-line drugs after the failure of

Biological Effects of HBV X Protein on Hepatocellular Carcinogenesis in Association with Cellular Factors

The X protein （HBx） of Human hepatitis B virus （HBV） acts as an indirect transcriptional transactivator to regulate the expression of many viral and cellular genes, as well as playing a critical role in pathogenesis and the deve!opment of Hepatocellular carcinoma （HCC）. Here we described the biological effects of HBx in association with four cellular factors, including inflammatory factors （COX-2 and iNOS）, oncoprotein （Ras）, and a newly identified tumor suppressor （YueF）. The characteristics of these effectors, which might be associated with hepatocellular carcinoma, are also discussed.

The mechanism of HBx protein to promote the initiation and progression of hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is the sixth most common malignancy worldwide and the third most common cause of death from cancer, after lung and stomach cancer. Hepatitis B virus(HBV) infection is closely related to HCC and is a major cause of HCC. HBV is a lysogenic virus of the hepadnavirus family. Its genome presents a slack, ring-like, double-chain structure, containing four open reading frames. The X region encodes the product HBV X protein(HBx), which is a multifunctional regulatory protein that plays an important role in intracellular signal transduction, viral genome replication and transcription, cell proliferation and apoptosis, cell cycle progression, protein degradation, and genetic stability of hepatocytes. This article summarizes the recent research on the mechanism of promotion of initiation and progression of HCC by HBx protein.

Hepatitis B Virus S Promoter Deletion in Hepatocellular Carcinoma

Objective To identify the difference and significance of dominant types of hepatitis B virus(HBV) pre-S mutation between liver tumor tissues and paired adjacent non-tumor tissues and to test if the mutations were tumor tissue specific.Methods HBV DNA isolated from 34 paired intratumoral and peritumoral tissues of Hepatocellular Carcinoma(HCC) patients were screened by PCR and direct sequencing. All patients carried HBV with genotype C, except for one B/C heterozygote. The expression, localization and excretion of LHBs mutant carrying pre-S deletions were characterized in vitro. The expression of endoplasmic reticulum(ER) GRP78 m RNA was assayed.Results Four patterns of pre-S mutations were identified: pre-S1 in-frame deletion involving the first start codon; pre-S2 in-frame deletion; pre-S2 start codon mutation with or without in-frame deletion; and S promoter in-frame deletion(ΔSP). The first two types were evenly found in both tumor and non-tumor tissues. They were rarely present as dominant strains. The last two types were frequently found in the dominant strains in tumor tissues. The overall prevalence of HBV carrying ΔSP was 17.64%(6/34) in tumor tissues, but none were dominant in non-tumor tissues. HBV carrying ΔSP was unable to produce S protein in vitro. Immunocytofluorescence assay showed that the ΔSP LHBs mutant aggregated in the cytoplasm, accumulating mainly in the ER. Transient transfection and expression of ΔSP mutant caused GRP78 up-regulation in vitro.Conclusion HBV S promoter deletion was found dominantly in HCC tumor tissue. The aggregation of mutant large surface proteins in the ER possibly involved in HBV-related HCC.

Hepatitis B Virus S Promoter Deletion in Hepatocellular Carcinoma

Objective To identify the difference and significance of dominant types of hepatitis B virus(HBV) pre-S mutation between liver tumor tissues and paired adjacent non-tumor tissues and to test if the mutations were tumor tissue specific.Methods HBV DNA isolated from 34 paired intratumoral and peritumoral tissues of Hepatocellular Carcinoma(HCC) patients were screened by PCR and direct sequencing. All patients carried HBV with genotype C, except for one B/C heterozygote. The expression, localization and excretion of LHBs mutant carrying pre-S deletions were characterized in vitro. The expression of endoplasmic reticulum(ER) GRP78 m RNA was assayed.Results Four patterns of pre-S mutations were identified: pre-S1 in-frame deletion involving the first start codon; pre-S2 in-frame deletion; pre-S2 start codon mutation with or without in-frame deletion; and S promoter in-frame deletion(ΔSP). The first two types were evenly found in both tumor and non-tumor tissues. They were rarely present as dominant strains. The last two types were frequently found in the dominant strains in tumor tissues. The overall prevalence of HBV carrying ΔSP was 17.64%(6/34) in tumor tissues, but none were dominant in non-tumor tissues. HBV carrying ΔSP was unable to produce S protein in vitro. Immunocytofluorescence assay showed that the ΔSP LHBs mutant aggregated in the cytoplasm, accumulating mainly in the ER. Transient transfection and expression of ΔSP mutant caused GRP78 up-regulation in vitro.Conclusion HBV S promoter deletion was found dominantly in HCC tumor tissue. The aggregation of mutant large surface proteins in the ER possibly involved in HBV-related HCC.