BACKGROUND: Hepatitis B virus (HBV) is an etiological factor of intrahepatic cholangiocarcinoma (ICC), but the pathogenic mechanisms remain unclear. This study aimed to investigate the expression and possible role of ...BACKGROUND: Hepatitis B virus (HBV) is an etiological factor of intrahepatic cholangiocarcinoma (ICC), but the pathogenic mechanisms remain unclear. This study aimed to investigate the expression and possible role of HBx, an HBV- encoded potentially oncogenic protein, in HBV-infected ICC. METHODS: Tissue samples were obtained from 54 specimens of HBV-infected ICC. Forty-four specimens were of peripheral type and 10 hilar type. Formalin-fixed, paraffin-embedded sections of the specimens were immunohistochemically stained for HBx and p53. RESULTS: HBx expression was found in 70.4% (38/54) of the specimens, and it was more frequently seen in the peripheral type than in the hilar type (79.5% vs 30.0%, P=0.002). All three well-differentiated ICCs expressed HBx, whereas 76.9% (30/39) moderately-differentiated and 41.7% (5/12) poorly-differentiated ICCs had HBx expression (P=0.033). Patients with HBx expression had a significantly higher prevalence of elevated serum alpha-fetoprotein (P=0.033). p53 protein expression was found in 18 of 54 cases (33.3%), and was not correlated with that of HBx. CONCLUSIONS: HBx may contribute to the pathogenesis of ICC, particularly the peripheral type. p53 abnormality may not play a significant role in HBx-mediated oncogenicity during ICC carcinogenesis.展开更多
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 H...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.展开更多
乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)对肝癌的发生发展具有十分重要的作用.HBx具有促进肝癌迁移的作用,但其作用的分子机制不清.本研究对HBx促进肝癌细胞迁移的分子机制进行了探讨.伤口愈合和Boyden’s chamber结果表明...乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)对肝癌的发生发展具有十分重要的作用.HBx具有促进肝癌迁移的作用,但其作用的分子机制不清.本研究对HBx促进肝癌细胞迁移的分子机制进行了探讨.伤口愈合和Boyden’s chamber结果表明,HBx可明显促进肝癌Hep G2细胞迁移.在稳定转染HBx的Hep G2(Hep G2-X)细胞中转染HBx结合蛋白(hepatitis B X-interacting protein,HBXIP)的RNA干扰片段,可明显抑制HBx的促迁移作用.免疫组化和实时定量PCR结果表明,HBXIP在肝癌组织中显著高表达,并且与HBx表达成正相关.荧光素酶报告基因和免疫印迹结果表明,HBx显著增强HBXIP的启动子活性和蛋白质表达水平.应用HBx的RNA干扰处理Hep G2-X细胞,HBXIP的启动子活性和蛋白质表达水平明显下降.将HBXIP启动子区的c AMP效应元件结合因子(CREB)结合位点突变后,HBx上调HBXIP的作用消失.应用CREB的RNA干扰处理肝癌细胞,在启动子水平和蛋白质水平上,HBx对HBXIP的上调作用被显著抑制.染色质免疫共沉淀结果表明,HBx能够通过CREB结合到HBXIP的启动子上,进而发挥激活HBXIP的功能.本研究结果表明,HBx促进肝癌细胞迁移的作用是通过CREB上调HBXIP实现的.这一发现对进一步揭示HBx促进肝癌细胞迁移的分子机制具有重要意义.展开更多
The chronic infection of hepatitis B virus(HBV) is closely related to the occurrence and development of hepatocellular carcinoma(HCC). Accumulated evidence has shown that HBV X protein(HBx protein) is a multifunctiona...The chronic infection of hepatitis B virus(HBV) is closely related to the occurrence and development of hepatocellular carcinoma(HCC). Accumulated evidence has shown that HBV X protein(HBx protein) is a multifunctional regulator with a crucial role in hepatocarcinogenesis. However, information on the mechanism by which HBV induces HCC is lacking. This review focuses on the pathological functions of HBx in HBV-induced hepatocarcinogenesis. As a transactivator, HBx can modulate nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB) and transcription factor AP-2. Moreover, HBx can affect regulatory non-coding RNAs(ncRNAs) including microRNAs and long ncRNAs(lncRNAs), such as miRNA-205 and highly upregulated in liver cancer(HULC), respectively. HBx is also involved in epigenetic modification, including methylation and acetylation. HBx interacts with various signal-transduction pathways, such as protein kinase B/Akt, Wnt/β-catenin, signal transducer and activator of transcription, and NF-κB pathways. Moreover, HBx affects cellular fate by shifting the balance toward cell survival. HBx may lead to the loss of apoptotic functions or directly contributes to oncogenesis by achieving transforming functions, which induce hepatocarcinogenesis. Additionally, HBx can modulate apoptosis and immune response by direct or indirect interaction with host factors. We conclude that HBx hastens the development of hepatoma.展开更多
Hepatitis B virus X(HBx)protein plays a pivotal role in the development of hepatitis B virus(HBV)-associated hepatocellular carcinoma.Although regulation of cytosolic calcium is essential for HBV replication and is me...Hepatitis B virus X(HBx)protein plays a pivotal role in the development of hepatitis B virus(HBV)-associated hepatocellular carcinoma.Although regulation of cytosolic calcium is essential for HBV replication and is mediated by HBx protein,the mechanism of HBx protein regulating intracellular calcium level remains poorly understood.The present study examined whether HBx protein elevated the intracellular calcium through interacting with storeoperated calcium entry(SOCE)components,Orai1 and stromal interaction molecule 1,and then identified the targets of HBx protein,with an attempt to understand the mechanism of HBx protein upsetting intracellular calcium homeostasis.By employing co-immunoprecipitation and GST-pull-down assay,we found that Orai1 protein interacted with HBx protein,and the C-terminus of Orai1 was implicated in the interaction.Confocal microscopy also revealed that HBx protein could co-localize with full-length Orai1 protein in HEK293 cells.Moreover,live cell calcium imaging exhibited that HBx protein elevated intracellular calcium,possibly by binding to SOCE components.Our results suggest that HBx protein binds to STIM1-Orai1 complexes to positively regulate the activity of plasma membrane store-operated calcium channels.展开更多
以PubMed和Web of Science数据库作为检索工具,采用文献计量学方法对乙型肝炎病毒X(hepatitis B virus X,HBx)研究文献从时间分布、期刊分布、国家分布等方面进行分析,通过可视化软件HistCite和CiteSpace对HBx相关文献生成可视化引文编...以PubMed和Web of Science数据库作为检索工具,采用文献计量学方法对乙型肝炎病毒X(hepatitis B virus X,HBx)研究文献从时间分布、期刊分布、国家分布等方面进行分析,通过可视化软件HistCite和CiteSpace对HBx相关文献生成可视化引文编年图及热点知识图谱,探讨HBx研究热点和前沿,揭示其发展历程。展开更多
目的构建靶向作用于小鼠肝前体细胞的动物模型,研究乙型肝炎病毒X蛋白(HBx)对肝前体细胞上皮间质转化(EMT)的影响。方法每周2次给予昆明小鼠2 m L/L四氯化碳灌胃,4周后行经门静脉注射稳定表达HBx的肝前体细胞和稳定表达空载体的肝前体...目的构建靶向作用于小鼠肝前体细胞的动物模型,研究乙型肝炎病毒X蛋白(HBx)对肝前体细胞上皮间质转化(EMT)的影响。方法每周2次给予昆明小鼠2 m L/L四氯化碳灌胃,4周后行经门静脉注射稳定表达HBx的肝前体细胞和稳定表达空载体的肝前体细胞同时切除部分肝脏。术后继续每周2次进行灌胃,并分别于术后3、5、7、14、21、28 d,处死小鼠取肝脏标本;实时定量PCR检测小鼠肝组织HBx的mRNA水平、免疫组织化学染色检测肝组织中外源性细胞的存活。实时荧光定量PCR检测上皮钙黏素(E-cadherin)、神经钙黏素(N-cadherin)、波形蛋白(vimentin)和细胞角蛋白18(CK18)mRNA水平,Western blot法检测E-cadherin、N-cadherin、vimentin、CK18蛋白水平。结果免疫组织化学染色结果显示,术后肝脏组织明显有外源性细胞存活.随注射时间延长,肝组织HBx的含量增加,表达区域增大;实时荧光定量PCR和Western blot结果均显示过表达HBx能够使小鼠肝组织中E-cadherin、CK18水平降低,而N-cadherin、vimentin的水平增加。结论动物模型证实HBx在肝前体细胞EMT过程中起着重要的调控作用。展开更多
目的:研究乙型肝炎病毒x基因(hepatitis B virus x gene,HBx)蛋白调节细胞内钙离子可能分子机制,揭示乙型肝炎病毒(hepatitis B virus,HBV)诱导肝癌的可能途径.方法:培养HEK293细胞,取第2代HEK293细胞转染pcDNA-HBx质粒,培养12、24和48 ...目的:研究乙型肝炎病毒x基因(hepatitis B virus x gene,HBx)蛋白调节细胞内钙离子可能分子机制,揭示乙型肝炎病毒(hepatitis B virus,HBV)诱导肝癌的可能途径.方法:培养HEK293细胞,取第2代HEK293细胞转染pcDNA-HBx质粒,培养12、24和48 h后,细胞活力细胞毒性检测(cell counting kit-8,CCK-8)观察转染后细胞生长情况,Western b l o t检测H B x蛋白的表达情况,当共同转染HBx基因、Orai1基因或STIM1基因后co-IP实验、免疫荧光检测观察细胞内蛋白结合情况.结果:转染pcDNA-HBx质粒后HBx蛋白可以在HEK293细胞高表达,转染后的24 h后细胞增殖加快(P<0.05),co-IP实验及免疫荧光检测结果均显示,HBx蛋白在细胞内可以与Orai1蛋白结合.结论:HBx蛋白通过与细胞膜钙离子通道Orai1结合,来升高细胞内钙离子浓度,从而影响细胞增殖等活性.展开更多
文摘BACKGROUND: Hepatitis B virus (HBV) is an etiological factor of intrahepatic cholangiocarcinoma (ICC), but the pathogenic mechanisms remain unclear. This study aimed to investigate the expression and possible role of HBx, an HBV- encoded potentially oncogenic protein, in HBV-infected ICC. METHODS: Tissue samples were obtained from 54 specimens of HBV-infected ICC. Forty-four specimens were of peripheral type and 10 hilar type. Formalin-fixed, paraffin-embedded sections of the specimens were immunohistochemically stained for HBx and p53. RESULTS: HBx expression was found in 70.4% (38/54) of the specimens, and it was more frequently seen in the peripheral type than in the hilar type (79.5% vs 30.0%, P=0.002). All three well-differentiated ICCs expressed HBx, whereas 76.9% (30/39) moderately-differentiated and 41.7% (5/12) poorly-differentiated ICCs had HBx expression (P=0.033). Patients with HBx expression had a significantly higher prevalence of elevated serum alpha-fetoprotein (P=0.033). p53 protein expression was found in 18 of 54 cases (33.3%), and was not correlated with that of HBx. CONCLUSIONS: HBx may contribute to the pathogenesis of ICC, particularly the peripheral type. p53 abnormality may not play a significant role in HBx-mediated oncogenicity during ICC carcinogenesis.
基金supported by the Program for Young Talents of Science and Technology in Universities of Inner Mongolia Autonomous Region NJYT-17-B30
文摘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.
文摘乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)对肝癌的发生发展具有十分重要的作用.HBx具有促进肝癌迁移的作用,但其作用的分子机制不清.本研究对HBx促进肝癌细胞迁移的分子机制进行了探讨.伤口愈合和Boyden’s chamber结果表明,HBx可明显促进肝癌Hep G2细胞迁移.在稳定转染HBx的Hep G2(Hep G2-X)细胞中转染HBx结合蛋白(hepatitis B X-interacting protein,HBXIP)的RNA干扰片段,可明显抑制HBx的促迁移作用.免疫组化和实时定量PCR结果表明,HBXIP在肝癌组织中显著高表达,并且与HBx表达成正相关.荧光素酶报告基因和免疫印迹结果表明,HBx显著增强HBXIP的启动子活性和蛋白质表达水平.应用HBx的RNA干扰处理Hep G2-X细胞,HBXIP的启动子活性和蛋白质表达水平明显下降.将HBXIP启动子区的c AMP效应元件结合因子(CREB)结合位点突变后,HBx上调HBXIP的作用消失.应用CREB的RNA干扰处理肝癌细胞,在启动子水平和蛋白质水平上,HBx对HBXIP的上调作用被显著抑制.染色质免疫共沉淀结果表明,HBx能够通过CREB结合到HBXIP的启动子上,进而发挥激活HBXIP的功能.本研究结果表明,HBx促进肝癌细胞迁移的作用是通过CREB上调HBXIP实现的.这一发现对进一步揭示HBx促进肝癌细胞迁移的分子机制具有重要意义.
文摘The chronic infection of hepatitis B virus(HBV) is closely related to the occurrence and development of hepatocellular carcinoma(HCC). Accumulated evidence has shown that HBV X protein(HBx protein) is a multifunctional regulator with a crucial role in hepatocarcinogenesis. However, information on the mechanism by which HBV induces HCC is lacking. This review focuses on the pathological functions of HBx in HBV-induced hepatocarcinogenesis. As a transactivator, HBx can modulate nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB) and transcription factor AP-2. Moreover, HBx can affect regulatory non-coding RNAs(ncRNAs) including microRNAs and long ncRNAs(lncRNAs), such as miRNA-205 and highly upregulated in liver cancer(HULC), respectively. HBx is also involved in epigenetic modification, including methylation and acetylation. HBx interacts with various signal-transduction pathways, such as protein kinase B/Akt, Wnt/β-catenin, signal transducer and activator of transcription, and NF-κB pathways. Moreover, HBx affects cellular fate by shifting the balance toward cell survival. HBx may lead to the loss of apoptotic functions or directly contributes to oncogenesis by achieving transforming functions, which induce hepatocarcinogenesis. Additionally, HBx can modulate apoptosis and immune response by direct or indirect interaction with host factors. We conclude that HBx hastens the development of hepatoma.
基金supported by grants from the National Natural Science Foundation of China(No.81001063)the Fundamental Research Funds for the Central Universities(No.2015QN150)
文摘Hepatitis B virus X(HBx)protein plays a pivotal role in the development of hepatitis B virus(HBV)-associated hepatocellular carcinoma.Although regulation of cytosolic calcium is essential for HBV replication and is mediated by HBx protein,the mechanism of HBx protein regulating intracellular calcium level remains poorly understood.The present study examined whether HBx protein elevated the intracellular calcium through interacting with storeoperated calcium entry(SOCE)components,Orai1 and stromal interaction molecule 1,and then identified the targets of HBx protein,with an attempt to understand the mechanism of HBx protein upsetting intracellular calcium homeostasis.By employing co-immunoprecipitation and GST-pull-down assay,we found that Orai1 protein interacted with HBx protein,and the C-terminus of Orai1 was implicated in the interaction.Confocal microscopy also revealed that HBx protein could co-localize with full-length Orai1 protein in HEK293 cells.Moreover,live cell calcium imaging exhibited that HBx protein elevated intracellular calcium,possibly by binding to SOCE components.Our results suggest that HBx protein binds to STIM1-Orai1 complexes to positively regulate the activity of plasma membrane store-operated calcium channels.
文摘以PubMed和Web of Science数据库作为检索工具,采用文献计量学方法对乙型肝炎病毒X(hepatitis B virus X,HBx)研究文献从时间分布、期刊分布、国家分布等方面进行分析,通过可视化软件HistCite和CiteSpace对HBx相关文献生成可视化引文编年图及热点知识图谱,探讨HBx研究热点和前沿,揭示其发展历程。
文摘目的:研究乙型肝炎病毒x基因(hepatitis B virus x gene,HBx)蛋白调节细胞内钙离子可能分子机制,揭示乙型肝炎病毒(hepatitis B virus,HBV)诱导肝癌的可能途径.方法:培养HEK293细胞,取第2代HEK293细胞转染pcDNA-HBx质粒,培养12、24和48 h后,细胞活力细胞毒性检测(cell counting kit-8,CCK-8)观察转染后细胞生长情况,Western b l o t检测H B x蛋白的表达情况,当共同转染HBx基因、Orai1基因或STIM1基因后co-IP实验、免疫荧光检测观察细胞内蛋白结合情况.结果:转染pcDNA-HBx质粒后HBx蛋白可以在HEK293细胞高表达,转染后的24 h后细胞增殖加快(P<0.05),co-IP实验及免疫荧光检测结果均显示,HBx蛋白在细胞内可以与Orai1蛋白结合.结论:HBx蛋白通过与细胞膜钙离子通道Orai1结合,来升高细胞内钙离子浓度,从而影响细胞增殖等活性.
