Clonal origin of hepatitis B virus-associated hepatocellular carcinoma

The clonal origin of tumors was studied in two multifocal hepatocellular carcinomas arising from hepatitis B viral cirrhosis. DNA extracted from several tumor nodules was analyzed for the presence of hepatitis B virus genome by the Southern blot technique. Unique clonal integration of the viral DNA sequences occurred in both cases. In each case, identical integration bands occurred among multiple nodules of liver tumor. These results are strong evidence of a unicentric origin of these tumors, although the tumors' gross appearance is suggestive of multifocal origin.     

Transgenic mouse models of hepatitis B virus-associated hepatocellular carcinoma

The multi-factorial and multi-step nature of cancer development makes analysis difficult in cell culture and non-genetic animal models. Recent progress in technology has allowed the development of several transgenic animal models addressing various aspects of liver diseases caused by hepatitis B virus in human patients. The experimental data from these studies in vivo highlight the importance of HBV gene products that alone or in conjunction with other host cellular protein(s) can deregulate the cell cycle control checkpoints in the hepatocytes of transgenic mice leading to the development of hepatocellular carcinoma. Moreover, these models are extremely useful in analysing and ascertaining the stages of malignant transformation linked to multiple genetic and non-genetic events of cancer development and in developing novel strategies of intervention.     

Occurrence of c-kit+ tumor cells in hepatitis B virus-associated hepatocellular carcinoma

Progenitor cells, termed oval cells, are involved in the pathogenesis of hepatocellular carcinoma (HCC) in animal models. By immunolabeling for c-kit and CD34 in human hepatitis B virus-associated cirrhosis with HCC (50 cases) and those with cirrhosis alone (10 cases), we found c-kit+ tumor cells in tumor tissue in 40 of 50 HCCs. The proportion was less than 0.1% of total tumor cell volume in most HCCs. Immunostaining for c-kit also was detected in sinusoidal endothelial cells in 43 of 50 HCCs. The incidence of oval cell occurrence in the adjacent nonneoplastic tissue in cases of HCC was high (44/50). The occurrence of oval cells, c-kit+ tumor cells, and c-kit+ sinusoidal cells in cases of human hepatitis B virus-associated HCC suggests that oval cell proliferation might be associated with the development of human hepatitis B virus-associated HCC. Furthermore, the c-kit+ sinusoidal cells might have a role in angiogenesis and progression of human hepatitis B virus-associated HCC.     

Detection of hepatitis C viraemia in Caucasian patients with hepatocellular carcinoma.

Potential risk factors for the development of hepatocellular carcinoma were analysed in 40 Caucasian patients with this malignancy. A higher proportion (14 of 40; 35%) had evidence of hepatitis C virus (HCV) infection than had evidence of either hepatitis B virus (HBV) carriage (17.5%) or alcohol abuse (30%). In all 14 patients whose sera were reactive by HCV ELISA (Ortho second generation test), the presence of antibodies to HCV were confirmed by recombinant immunoblot assay (Ortho RIBA-2). Furthermore, two independent laboratories detected HCV-RNA in 10 of the 14 (71%) anti-HCV positive sera. Two additional sera were shown to contain HCV-RNA when reanalysed by a modified PCR using oligonucleotide primers designed to amplify a shorter fragment of the 5' noncoding region of the genome. Seven of the anti-HCV positive patients also had evidence of prior HBV infection and 2 admitted to alcohol abuse. HCV infection was the only identifiable risk factor in 6 patients. These data confirm the association between HCV infection and hepatocellular carcinoma and suggest that persistent viral replication accompanies tumour development in the majority of patients whose serum contains anti-HCV.     

Serum antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma

Antibodies against hepatitis C virus (anti-HCV) were detected in 60.8% of 78 patients with hepatocellular carcinoma (HCC). Cirrhosis, present in most of the patients, as well as alcohol abuse, age, sex, and alpha-fetoprotein were equally distributed in the anti-HCV-positive and -negative groups. HBsAg positivity was significatively higher in negative anti-HCV group. By contrast, hepatitis B virus (HBV) antibodies were detected more frequently in positive anti-HCV patients than in the negative anti-HCV group. These data must be considered with caution because of the small number of HBsAg-positive patients. It is concluded that the high prevalence of anti-HCV in patients with HCC may suggest an etiological role of the hepatitis C virus, although in relationship to age, alcohol abuse and cirrhosis, the similarity in the two groups questions this hypothesis.     

Direct-acting antivirals for patients with chronic hepatitis C and hepatocellular carcinoma in Taiwan

Background/purposeThe treatment of chronic hepatitis C (CHC) has evolved from interferon (IFN)-based therapy to direct acting antivirals (DAAs). The effect of antiviral treatment on outcome of hepatocellular carcinoma (HCC) patients with CHC has not been well analyzed in Taiwan.MethodsFrom April 2015 to May 2018, 199 HCC patients with CHC undergoing DAAs treatment, including 127 having prospectively longitudinal observation, were enrolled. Among them, 107 BCLC 0/A patients achieving curative treatment of HCC were further compared with a historical cohort of 42 HCC patients experienced pegylated interferon (Peg-IFN) plus ribavirin for CHC after curative treatment.ResultsThe sustained virological response (SVR) rates were 95.0% in BCLC stage 0/A (114/120), 97.1% in BCLC B (68/70), and 77.8% in BCLC C (7/9). The median recurrence-free survivals (RFS) between the DAA and IFN arms were of no difference by counting either from antiviral treatment (29.3 mo vs 39.2 mo, p = 0.764) or from curative treatment (65.8 mo vs 44.0 mo, p = 0.130), respectively. Achievement of SVR was the key independent factor associated with RFS and overall survival. The pattern of recurrence was also similar between the DAA and IFN arms. For intermediate stage HCC patients, the median time to tumor progression was 9.2 months from the initiation of DAA therapy, and 90% of patients maintained in BCLC B till 12 months after the DAA treatment.ConclusionsThe SVR is high within BCLC B HCV-HCC patients by DAAs treatment. The risk of HCC recurrence and progression is not increased by DAAs.     

RIPK3 mRNA level acts as a diagnostic biomarker in hepatitis B virus-associated hepatocellular carcinoma

HBV-associated hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death worldwide, and non-invasive early detection of HBV-associated HCC requires to be improved. To determine the alteration and clinical relevance of necroptosis and its key regulator receptor-interacting protein kinase 3 (RIPK3) in HBV-associated HCC, we detected the mRNA level of RIPK3 in peripheral blood mononuclear cells (PBMCs) and analyzed its correlation with clinical parameters. Here, we demonstrate that the expression of RIPK3 is elevated in patients with HBV-associated HCC compared to patients with chronic hepatitis B (CHB) and patients with HBV-related liver cirrhosis (LC). The mRNA level of RIPK3 is positively correlated with the severity of clinical manifestations and TNM stages. Moreover, the serum levels of RIPK3-asssocited cytokines are altered in consistent with the change of RIPK3 expression. The diagnostic accuracy of RIPK3 mRNA level is comparable to AFP test in discriminating HBV-associated HCC from LC and is better than AFP test in discriminating HBV-associated HCC from CHB. The combination of RIPK3 mRNA level and AFP test significantly improves the diagnosis of HBV-associated HCC. These data suggest that RIPK3 mRNA level is a biomarker in the onset and progression of HBV-associated HCC and may provide novel diagnostic strategies combined with the AFP test.     

Tumor suppressor genes,growth factor genes,and oncogenes in hepatitis B virus-associated hepatocellular carcinoma

A series of changes in the genes that control hepatocyte growth, or interference with the protein products of these genes, appears to have an important role in the etiology of hepatocellular carcinoma (HCC). Mutations of the p53 tumor suppressor gene have been identified in 30-50% of HCC patients in some geographic areas. Abnormalities of the RB tumor suppressor gene have been found in 20-25% of HCCs, including 80-86% of HCCs with p53 mutations. Overexpression of transforming growth factor α (TGF-a), insulin-like growth factor II (IGF-II), and the oncogenes N-ras, c-myc, and c-fos have been found in high percentages of HCC patients. The cumulative effect of these changes may be more important than the order in which they occur. Some of these changes may explain the mechanism(s) by which the hepatitis B virus participates in the development of HCC. © 1994 Wiley-Liss, Inc.     

Plasma myeloperoxidase levels correlate with hepatocellular carcinoma in chronic hepatitis C

Myeloperoxidase (MPO) is an enzyme responsible for generating hypochlorous acid and reactive oxidants that may lead to liver injury and cancer in hepatitis C (HCV) infection. MPO expression level is regulated by a polymorphism in the promoter region −463 of MPO gene. In the current study, MPO plasma levels and the G-463A MPO polymorphism were determined in 158 chronically HCV infected patients with and without hepatocellular carcinoma (HCC). MPO plasma levels were determined using a commercially ELISA kit. The G-463A MPO polymorphism was accessed by real time PCR using TaqMan probes. The MPO plasma levels of patients with HCV-HCC were higher in comparison to patients with chronic hepatitis or with those patients with severe fibrosis (p = 0.01 and p = 0.04, respectively). The MPO G-463A polymorphism was not associated with HCV outcome. These findings suggest MPO levels monitoring may be a potential biological marker to HCC screening in patients with HCV.     

具有丙型肝炎病毒抗原标志的肝细胞癌的病理学特征

目的系统地分析具有不同肝炎病毒抗原标志的肝细胞癌（ＨＣＣ）的组织病理学特征。方法用免疫组化ＰＡＰ法、ＡＢＣ法显示６９例组织中ＨＢＶ及ＨＣＶ抗原，运用ＣＡＳ－２００系统对癌组织进行图像分析。结果与单独ＨＢＶ感染者相比，单独ＨＣＶ感染相关ＨＣＣ中透明细胞型肝癌发生率较高（７／９比４／３３），分化较好；癌周肝窦内和汇管区淋巴细胞浸润及肝细胞坏死较轻（Ｐ＜０．０１）；小胆管破坏较常见，且与淋巴滤泡样结构形成关系密切（Ｐ＜０．０５）。此外，该组患者年龄较大，临床症状较轻，手术预后较佳。结论单独ＨＣＶ感染相关ＨＣＣ具有不同于单独ＨＢＶ感染相关ＨＣＣ的临床及病理学特征，癌细胞核ＤＮＡ含量及形态学参数定量分析结果与其生物学行为相符。     

Clinicopathologic and prognostic significance of the histologic activity of noncancerous liver tissue in hepatitis B virus-associated hepatocellular carcinoma

We prospectively studied 66 patients infected with the hepatitis B virus who underwent liver resection for hepatocellular carcinoma (HCC) to evaluate the influence of the histologic activity of noncancerous liver tissue on clinicopathologic features and prognosis. Based on the histologic activity index (HAI) score of nontumorous liver tissue, patients were classified into 3 groups: mild, moderate, or severe hepatitis. Overall, higher HAI scores were more frequent in patients with poorer liver function: lower serum albumin levels and higher indocyanine green retention at 15 minutes. Moreover, patients with moderate hepatitis had more frequent venous invasion, and the tumor size decreased with increasing HAI scores. Similar results were observed when the fibrosis category was excluded in the calculation of HAI scores. The overall or disease-free survival rates did not differ significantly among the 3 groups of patients. However, higher fibrosis scores were associated significantly with shorter disease-free survival rates. HAI scores correlated significantly with certain clinicopathologic features. In patients with hepatitis B-related HCC, a higher fibrosis score in the nontumorous liver tissue, but not histologic hepatitic activity, seems to be a significant factor predisposing to shorter survival.     

Frequent detection of hepatitis B virus DNA in hepatocellular carcinoma of patients with sustained virologic response for hepatitis C virus

Hepatocellular carcinoma (HCC) develops several years after the eradication of hepatitis C virus (HCV) by interferon therapy. Risk factors for the development of HCC are only partly understood. To elucidate the role of occult hepatitis B virus (HBV) infection in hepatocarcinogenesis in patients with sustained virologic response, the prevalences of HBV‐related makers were examined. Study group comprised 16 patients with sustained virologic response (group A) and 50 with HCV (group B). Anti‐HBc and anti‐HBs in serum were examined by enzyme‐linked immunoassay. HBV DNA in liver was examined by nested polymerase chain reaction, using primers specific for genes encoding for HBx, HBsAg, HBcAg, and HBV cccDNA. Sequence of the amplified HBV DNA for ‘a’ determinant of HBsAg was determined in HCC. Anti‐HBc was positive in 10 of 16 in group A and 25 of 50 in group B. HBV DNA in liver was detected in 12 of 16 in group A and 21 of 50 in group B (P = 0.044). In group A, HBV DNA in liver was detected frequently in patients without cirrhosis and in those with a longer period from the time of HCV eradication to the development of HCC. Mutation in ‘a’ determinant of HBsAg was found in three HCC of group A. Occult HBV infection may be one of the most important risk factors in hepatocarcinogenesis of Japanese patients with sustained virologic response. J. Med. Virol. 81:1009–1014, 2009. © 2009 Wiley‐Liss, Inc.     

Chronic hepatitis C virus infection and the pathogenesis of hepatocellular carcinoma

There is a strong epidemiologic relationship between chronic hepatitis C virus (HCV) infection and the development of hepatocellular carcinoma, although the cellular and molecular mechanisms of tumor formation remain to be firmly established. Clearly, HCV is associated with the development of chronic hepatitis and cirrhosis, so it may contribute to hepatocarcinogenesis as a consequence of its central role in the appearance and progression of necroinflammatory liver disease. There is also increasing evidence for a direct contribution of several HCV gene products to the development of the transformed phenotype, although none of the putative mechanisms involved in tumor formation have been strongly supported by in vivo evidence. Even if HCV is not shown to be a complete carcinogen, it may act as a co-carcinogen with underlying (serologically negative) hepatitis B virus infection, in the context of alcoholic cirrhosis, and in patients with long term exposure to chemical hepatocarcinogens such as aflatoxin B1.     

丙型肝炎病毒相关性肝细胞癌的基因表达分析

目的分析慢性丙型肝炎发展为肝细胞癌(HCC)过程中的差异表达基因谱。方法以GEO数据库的基因表达谱数据GDS4880、GDS4887为分析材料,采用Qlucore Omics Explorer 3.0软件筛选慢性丙型肝炎与丙型肝炎病毒(HCV)相关性肝细胞癌芯片数据的差异表达基因,结合生物信息学工具PANTHER、DAVID、STRING、Cytoscape对差异表达基因及其相互作用关系进行分析。结果筛选出共差异表达基因328个,其中上调表达133个,下调表达195个。这些差异表达基因主要涉及代谢过程、生物调节、定位等生物过程,p53信号通路、胰岛素信号通路、磷脂酰肌醇信号系统、细胞凋亡等信号通路。差异表达基因编码蛋白间的相互作用主要集中在14个蛋白质(CYP2B6、CYP2E1、AKT1、CDK16、RELA、CDC27、PIK3CA、GNB1、FOXO1、FYN、PDPK1、SCAND1、SGOL1、RPH3AL)。结论 CYP2E1等14个基因可能与HCV相关性肝细胞癌的发生发展相关。