The Experts below are selected from a list of 3297 Experts worldwide ranked by ideXlab platform
Xiaohua Jiang - One of the best experts on this subject based on the ideXlab platform.
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway.
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P
-
effects of hepatitis c virus core Protein and Nonstructural Protein 4B on the wnt β catenin pathway
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P < 0.01), and promoted the nuclear translocation of β-catenin (F = 66.54, P < 0.01) and the Wnt3a-induced proliferation of LO2 cells (P < 0.01 or P < 0.05). Moreover, activation of the Wnt/β-catenin signaling pathway was greater with the core Protein than with NS4B (P < 0.01 or P < 0.05). HCV core Protein and NS4B directly activate the Wnt/β-catenin signaling pathway in Huh7 cells and LO2 cells induced by Wnt3a. These data suggest that HCV core Protein and NS4B contribute to HCV-associated hepatocellular carcinogenesis.
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway
BMC, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Abstract Background Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Results Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P
-
Effect of hepatitis C virus core Protein and NS4B on the proliferation of HepG2 cells
Chinese Journal of Clinical Hepatology, 2014Co-Authors: Xiaohua JiangAbstract:Objective To investigate the effect of hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B(NS4B) on the proliferation of HepG2 cells and its possible mechanism.Methods The two recombinant plasmid pcDNA3.1 (-)Core and pcDNA3.1 (-) NS4B were transiently transfected respectively and co-transfected into HepG2 cells by lipofectamine, simultaneously HepG2 cells transfected with pcDNA3.1 (-) and untransfected HepG2 cells were used as control.The expression of mRNA and Protein of HCV Core,NS4B,Wnt1,β-catenin,c-myc and CyclinDl in the cells of each group were detected by RT-PCR and Western blot respectively.Cell proliferation changes were measured by MTT assay and plate colony formation assay.The cell cycle distribution was tested by flow cytometry.Results ①HCV Core or/and NS4B mRNA and Protein were expressed successfully in the HepG2 cells transfected with pcDNA3.1 (-)Core,pcDNA3.1 (-)NS4B alone or in combination.② The relative expression levels of mRNA and Protein of Wnt1,β-catenin,c-myc and CyclinD1 were higher in the cells transfected with pcDNA3.1 (-) Core,pcDNA3.1 (-) NS4B alone or in combination than those in the cells transfected with pcDNA3.1 (-) and untransfected HepG2 cells(P ＜0.01).③Compared with the HepG2 cells transfected with pcDNA3.1 (-)and untransfected,cell viability,cloning efficiency and the percentage of cells at S and G2/M phases were significantly increased in the cells transfected with pcDNA3.1 (-) Core,pcDNA3.1 (-) NS4B alone or in combination (P ＜ 0.01).Conclusion HCV core Protein and NS4B can accelerate cell cycle progression and promote cell proliferation of HepG2 cells probably through enhancement of Wntl,β-catenin,c-myc and CyclinD1 expression. Key words: Hepatitis viruses; Viral core Proteins; Viral Nonstructural Proteins; Cell division
-
screening of differentially expressed genes and gene pathways in hepatitis c virus 1b type Nonstructural Protein 4B stably expressed l02 cell line
Journal of Central South University. Medical sciences, 2014Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Chuang Lei, Bo Jiang, Hua PengAbstract:OBJECTIVE To screen differentially expressed genes and gene pathways in L02 cell line stably expressing hepatitis C virus (HCV) Ib type Nonstructural Protein 4B (NS4B) mediated by lentiviral system, and to provide a basis for further research of molecular biological mechanism of NS4B gene in chronic hepatitis C and hepatocarcinogenesis. METHODS NS4B stably overexpressed L02 cell line and negative control stable L02 cell line, designated as L02-NS4B and L02-mkate2 respectively, were resurrected and amplified in vitro. The differentially expressed genes between L02-NS4B and L02-mkate2 were determined by gene expression microarray from Human Gene 1.0ST. The significant pathways of the differential genes were selected by the Fisher's exact test and χ2 test according to kyoto encyclopedia of genes and genomes (KEGG) database. The differential expression levels of 5 selected genes including Protein kinase C delta binding Protein (PRKCDBP), tumor Protein p53 (TP53), v-akt murine thymoma viral oncogene homolog 1 (AKT1), baculoviral IAP repeat containing 3 (BIRC3) and B-cell lymphoma 2-like1 (BCL2L1) from cDNA microarray data were further verified by real-time quantitative polymerase chain reaction (real-time QPCR). RESULTS Between L02-NS4B and L02-mkate2, the genes with flurescence intensity ratio >1.2 or <0.8 were considered as differentially expressed genes. A total of 2 682 differentially expressed genes in the known 28 869 human genes were detected in L02-NS4B, 1 446 genes were upregulated and 1 236 genes were downregulated. A total of 41 involved pathways of up-regulated differential genes were identified by KEGG database, mainly including apoptosis, extracellular matrix receptor interaction, cell cycle, pathways in cancer and Toll-like receptor signaling pathway; and 20 involved pathways of down-regulated differential genes were identified, mainly including pathways in cancer, Wnt signaling pathway and cell cycle pathway. Of the 5 upregulated genes selected from cDNA microarray data, 3 genes showed the same differential expression pattern by real-time QPCR as that shown in cDNA microarray data, namely AKT1, BIRC3 and BCL2L1. The confirmation rate of real-time QPCR was 60%. CONCLUSION HCVNS4B can up-regulate or down-regulate the expression of many genes in L02 cells, thus affecting multiple signaling pathways relevant to cell apoptosis, cell cycle and carcinogenesis.
Ralf Bartenschlager - One of the best experts on this subject based on the ideXlab platform.
-
glycine zipper motifs in hepatitis c virus Nonstructural Protein 4B are required for the establishment of viral replication organelles
Journal of Virology, 2017Co-Authors: David L Paul, Vlastimil Jirasko, Vanesa Madan, Omar Ramirez, Maja Bencun, Ina Karen Stoeck, Ralf BartenschlagerAbstract:: Hepatitis C virus (HCV) RNA replication occurs in tight association with remodeled host cell membranes, presenting as cytoplasmic accumulations of single-, double-, and multimembrane vesicles in infected cells. Formation of these so-called replication organelles is mediated by a complex interplay of host cell factors and viral replicase Proteins. Of these, Nonstructural Protein 4B (NS4B), an integral transmembrane Protein, appears to play a key role, but little is known about the molecular mechanisms of how this Protein contributes to organelle biogenesis. Using forward and reverse genetics, we identified glycine zipper motifs within transmembrane helices 2 and 3 of NS4B that are critically involved in viral RNA replication. Foerster resonance energy transfer analysis revealed the importance of the glycine zippers in NS4B homo- and heterotypic self-interactions. Additionally, ultrastructural analysis using electron microscopy unraveled a prominent role of glycine zipper residues for the subcellular distribution and the morphology of HCV-induced double-membrane vesicles. Notably, loss-of-function NS4B glycine zipper mutants prominently induced single-membrane vesicles with secondary invaginations that might represent an arrested intermediate state in double-membrane vesicle formation. These findings highlight a so-far-unknown role of glycine residues within the membrane integral core domain for NS4B self-interaction and functional as well as structural integrity of HCV replication organelles.IMPORTANCE Remodeling of the cellular endomembrane system leading to the establishment of replication organelles is a hallmark of positive-strand RNA viruses. In the case of HCV, expression of the Nonstructural Proteins induces the accumulation of double-membrane vesicles that likely arise from a concerted action of viral and coopted cellular factors. However, the underlying molecular mechanisms are incompletely understood. Here, we identify glycine zipper motifs within HCV NS4B transmembrane segments 2 and 3 that are crucial for the Protein's self-interaction. Moreover, glycine residues within NS4B transmembrane helices critically contribute to the biogenesis of functional replication organelles and, thus, efficient viral RNA replication. These results reveal how glycine zipper motifs in NS4B contribute to structural and functional integrity of the HCV replication organelles and, thus, viral RNA replication.
-
a combined genetic proteomic approach identifies residues within dengue virus ns4B critical for interaction with ns3 and viral replication
Journal of Virology, 2015Co-Authors: Laurent Chatelchaix, Pietro Scaturro, Mirko Cortese, Stephanie Kallis, Marie Bartenschlager, Wolfgang Fischl, Bernd Fischer, Ralf BartenschlagerAbstract:UNLABELLED: Dengue virus (DENV) infection causes the most prevalent arthropod-borne viral disease worldwide. Approved vaccines are not available, and targets suitable for the development of antiviral drugs are lacking. One possible drug target is Nonstructural Protein 4B (NS4B), because it is absolutely required for virus replication; however, its exact role in the DENV replication cycle is largely unknown. With the aim of mapping NS4B determinants critical for DENV replication, we performed a reverse genetic screening of 33 NS4B mutants in the context of an infectious DENV genome. While the majority of these mutations were lethal, for several of them, we were able to select for second-site pseudoreversions, most often residing in NS4B and restoring replication competence. To identify all viral NS4B interaction partners, we engineered a fully viable DENV genome encoding an affinity-tagged NS4B. Mass spectrometry-based analysis of the NS4B complex isolated from infected cells identified the NS3 protease/helicase as a major interaction partner of NS4B. By combining the genetic complementation map of NS4B with a replication-independent expression system, we identified the NS4B cytosolic loop-more precisely, amino acid residue Q134-as a critical determinant for NS4B-NS3 interaction. An alanine substitution at this site completely abrogated the interaction and DENV RNA replication, and both were restored by pseudoreversions A69S and A137V. This strict correlation between the degree of NS4B-NS3 interaction and DENV replication provides strong evidence that this viral Protein complex plays a pivotal role during the DENV replication cycle, hence representing a promising target for novel antiviral strategies. IMPORTANCE: With no approved therapy or vaccine against dengue virus infection, the viral Nonstructural Protein 4B (NS4B) represents a possible drug target, because it is indispensable for virus replication. However, little is known about its precise structure and function. Here, we established the first comprehensive genetic interaction map of NS4B, identifying amino acid residues that are essential for virus replication, as well as second-site mutations compensating for their defects. Additionally, we determined the NS4B viral interactome in infected cells and identified the NS3 protease/helicase as a major interaction partner of NS4B. We mapped residues in the cytosolic loop of NS4B as critical determinants for interaction with NS3, as well as RNA replication. The strong correlation between NS3-NS4B interaction and RNA replication provides strong evidence that this complex plays a pivotal role in the viral replication cycle, hence representing a promising antiviral drug target.
-
The predominant species of Nonstructural Protein 4B in hepatitis C virus-replicating cells is not palmitoylated.
Journal of General Virology, 2015Co-Authors: David L Paul, Ralf Bartenschlager, Christopher MccormickAbstract:Hepatitis C virus (HCV) represents a significant global health burden. Viral replication is thought to occur in close association with remodelled host cell membranes, with non-structural Protein 4B (NS4B) being a key player in this process. NS4B is a poorly characterized integral membrane Protein, which has been reported to be palmitoylated at its carboxy-terminal end. In order to extend this observation and to establish a functional role for NS4B palmitoylation, we sought to determine the status of this post-translational modification when the Protein was expressed as part of a functional viral replicase. We performed direct metabolic labelling and polyethylene glycol-maleimide palmitoylation reporter assays on NS4B expressed in cells containing subgenomic replicons and infectious viral RNA. In a vaccinia virus-based expression system NS4B palmitoylation was detected in a genotype-dependent manner. However, in spite of the high sensitivity of the methods used, no NS4B palmitoylation was found in physiologically more relevant systems. Thus, NS4B palmitoylation is most likely dispensable for HCV RNA replication.
-
Aminoterminal amphipathic α-helix AH1 of hepatitis C virus Nonstructural Protein 4B possesses a dual role in RNA replication and virus production.
PLoS pathogens, 2014Co-Authors: Jerome Gouttenoire, Ralf Bartenschlager, Roland Montserret, François Penin, David Paul, Rosa Castillo, Simon Meister, Darius MoradpourAbstract:Nonstructural Protein 4B (NS4B) is a key organizer of hepatitis C virus (HCV) replication complex formation. In concert with other Nonstructural Proteins, it induces a specific membrane rearrangement, designated as membranous web, which serves as a scaffold for the HCV replicase. The N-terminal part of NS4B comprises a predicted and a structurally resolved amphipathic α-helix, designated as AH1 and AH2, respectively. Here, we report a detailed structure-function analysis of NS4B AH1. Circular dichroism and nuclear magnetic resonance structural analyses revealed that AH1 folds into an amphipathic α-helix extending from NS4B amino acid 4 to 32, with positively charged residues flanking the helix. These residues are conserved among hepaciviruses. Mutagenesis and selection of pseudorevertants revealed an important role of these residues in RNA replication by affecting the biogenesis of double-membrane vesicles making up the membranous web. Moreover, alanine substitution of conserved acidic residues on the hydrophilic side of the helix reduced infectivity without significantly affecting RNA replication, indicating that AH1 is also involved in virus production. Selective membrane permeabilization and immunofluorescence microscopy analyses of a functional replicon harboring an epitope tag between NS4B AH1 and AH2 revealed a dual membrane topology of the N-terminal part of NS4B during HCV RNA replication. Luminal translocation was unaffected by the mutations introduced into AH1, but was abrogated by mutations introduced into AH2. In conclusion, our study reports the three-dimensional structure of AH1 from HCV NS4B, and highlights the importance of positively charged amino acid residues flanking this amphipathic α-helix in membranous web formation and RNA replication. In addition, we demonstrate that AH1 possesses a dual role in RNA replication and virus production, potentially governed by different topologies of the N-terminal part of NS4B.
-
Morphological and biochemical characterization of the membranous hepatitis C virus replication compartment
Journal of Virology, 2013Co-Authors: David L Paul, Simone Hoppe, Gesine Saher, Jacomine Krijnse-locker, Ralf BartenschlagerAbstract:Like all other positive-strand RNA viruses, hepatitis C virus (HCV) induces rearrangements of intracellular membranes that are thought to serve as a scaffold for the assembly of the viral replicase machinery. The most prominent membranous structures present in HCV-infected cells are double-membrane vesicles (DMVs). However, their composition and role in the HCV replication cycle are poorly understood. To gain further insights into the biochemcial properties of HCV-induced membrane alterations, we generated a functional replicon containing a hemagglutinin (HA) affinity tag in Nonstructural Protein 4B (NS4B), the supposed scaffold Protein of the viral replication complex. By using HA-specific affinity purification we isolated NS4B-containing membranes from stable replicon cells. Complementing biochemical and electron microscopy analyses of purified membranes revealed predominantly DMVs, which contained viral Proteins NS3 and NS5A as well as enzymatically active viral replicase capable of de novo synthesis of HCV RNA. In addition to viral factors, co-opted cellular Proteins, such as vesicle-associated membrane Protein-associated Protein A (VAP-A) and VAP-B, that are crucial for viral RNA replication, as well as cholesterol, a major structural lipid of detergent-resistant membranes, are highly enriched in DMVs. Here we describe the first isolation and biochemical characterization of HCV-induced DMVs. The results obtained underline their central role in the HCV replication cycle and suggest that DMVs are sites of viral RNA replication. The experimental approach described here is a powerful tool to more precisely define the molecular composition of membranous replication factories induced by other positive-strand RNA viruses, such as picorna-, arteri- and coronaviruses.
Jing Ren - One of the best experts on this subject based on the ideXlab platform.
-
effects of hepatitis c virus core Protein and Nonstructural Protein 4B on the wnt β catenin pathway
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P < 0.01), and promoted the nuclear translocation of β-catenin (F = 66.54, P < 0.01) and the Wnt3a-induced proliferation of LO2 cells (P < 0.01 or P < 0.05). Moreover, activation of the Wnt/β-catenin signaling pathway was greater with the core Protein than with NS4B (P < 0.01 or P < 0.05). HCV core Protein and NS4B directly activate the Wnt/β-catenin signaling pathway in Huh7 cells and LO2 cells induced by Wnt3a. These data suggest that HCV core Protein and NS4B contribute to HCV-associated hepatocellular carcinogenesis.
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway.
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway
BMC, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Abstract Background Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Results Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P
Yu-tao Xie - One of the best experts on this subject based on the ideXlab platform.
-
effects of hepatitis c virus core Protein and Nonstructural Protein 4B on the wnt β catenin pathway
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P < 0.01), and promoted the nuclear translocation of β-catenin (F = 66.54, P < 0.01) and the Wnt3a-induced proliferation of LO2 cells (P < 0.01 or P < 0.05). Moreover, activation of the Wnt/β-catenin signaling pathway was greater with the core Protein than with NS4B (P < 0.01 or P < 0.05). HCV core Protein and NS4B directly activate the Wnt/β-catenin signaling pathway in Huh7 cells and LO2 cells induced by Wnt3a. These data suggest that HCV core Protein and NS4B contribute to HCV-associated hepatocellular carcinogenesis.
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway.
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway
BMC, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Abstract Background Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Results Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P
-
screening of differentially expressed genes and gene pathways in hepatitis c virus 1b type Nonstructural Protein 4B stably expressed l02 cell line
Journal of Central South University. Medical sciences, 2014Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Chuang Lei, Bo Jiang, Hua PengAbstract:OBJECTIVE To screen differentially expressed genes and gene pathways in L02 cell line stably expressing hepatitis C virus (HCV) Ib type Nonstructural Protein 4B (NS4B) mediated by lentiviral system, and to provide a basis for further research of molecular biological mechanism of NS4B gene in chronic hepatitis C and hepatocarcinogenesis. METHODS NS4B stably overexpressed L02 cell line and negative control stable L02 cell line, designated as L02-NS4B and L02-mkate2 respectively, were resurrected and amplified in vitro. The differentially expressed genes between L02-NS4B and L02-mkate2 were determined by gene expression microarray from Human Gene 1.0ST. The significant pathways of the differential genes were selected by the Fisher's exact test and χ2 test according to kyoto encyclopedia of genes and genomes (KEGG) database. The differential expression levels of 5 selected genes including Protein kinase C delta binding Protein (PRKCDBP), tumor Protein p53 (TP53), v-akt murine thymoma viral oncogene homolog 1 (AKT1), baculoviral IAP repeat containing 3 (BIRC3) and B-cell lymphoma 2-like1 (BCL2L1) from cDNA microarray data were further verified by real-time quantitative polymerase chain reaction (real-time QPCR). RESULTS Between L02-NS4B and L02-mkate2, the genes with flurescence intensity ratio >1.2 or <0.8 were considered as differentially expressed genes. A total of 2 682 differentially expressed genes in the known 28 869 human genes were detected in L02-NS4B, 1 446 genes were upregulated and 1 236 genes were downregulated. A total of 41 involved pathways of up-regulated differential genes were identified by KEGG database, mainly including apoptosis, extracellular matrix receptor interaction, cell cycle, pathways in cancer and Toll-like receptor signaling pathway; and 20 involved pathways of down-regulated differential genes were identified, mainly including pathways in cancer, Wnt signaling pathway and cell cycle pathway. Of the 5 upregulated genes selected from cDNA microarray data, 3 genes showed the same differential expression pattern by real-time QPCR as that shown in cDNA microarray data, namely AKT1, BIRC3 and BCL2L1. The confirmation rate of real-time QPCR was 60%. CONCLUSION HCVNS4B can up-regulate or down-regulate the expression of many genes in L02 cells, thus affecting multiple signaling pathways relevant to cell apoptosis, cell cycle and carcinogenesis.
-
Screening of differentially expressed genes and gene pathways in hepatitis C virus 1b type Nonstructural Protein 4B stably expressed L02 cell line
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences, 2014Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Chuang Lei, Bo Jiang, Hua PengAbstract:OBJECTIVE To screen differentially expressed genes and gene pathways in L02 cell line stably expressing hepatitis C virus (HCV) Ib type Nonstructural Protein 4B (NS4B) mediated by lentiviral system, and to provide a basis for further research of molecular biological mechanism of NS4B gene in chronic hepatitis C and hepatocarcinogenesis. METHODS NS4B stably overexpressed L02 cell line and negative control stable L02 cell line, designated as L02-NS4B and L02-mkate2 respectively, were resurrected and amplified in vitro. The differentially expressed genes between L02-NS4B and L02-mkate2 were determined by gene expression microarray from Human Gene 1.0ST. The significant pathways of the differential genes were selected by the Fisher's exact test and χ2 test according to kyoto encyclopedia of genes and genomes (KEGG) database. The differential expression levels of 5 selected genes including Protein kinase C delta binding Protein (PRKCDBP), tumor Protein p53 (TP53), v-akt murine thymoma viral oncogene homolog 1 (AKT1), baculoviral IAP repeat containing 3 (BIRC3) and B-cell lymphoma 2-like1 (BCL2L1) from cDNA microarray data were further verified by real-time quantitative polymerase chain reaction (real-time QPCR). RESULTS Between L02-NS4B and L02-mkate2, the genes with flurescence intensity ratio >1.2 or
Ya-ping Cai - One of the best experts on this subject based on the ideXlab platform.
-
effects of hepatitis c virus core Protein and Nonstructural Protein 4B on the wnt β catenin pathway
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P < 0.01), and promoted the nuclear translocation of β-catenin (F = 66.54, P < 0.01) and the Wnt3a-induced proliferation of LO2 cells (P < 0.01 or P < 0.05). Moreover, activation of the Wnt/β-catenin signaling pathway was greater with the core Protein than with NS4B (P < 0.01 or P < 0.05). HCV core Protein and NS4B directly activate the Wnt/β-catenin signaling pathway in Huh7 cells and LO2 cells induced by Wnt3a. These data suggest that HCV core Protein and NS4B contribute to HCV-associated hepatocellular carcinogenesis.
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway.
BMC Microbiology, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P
-
Effects of hepatitis C virus core Protein and Nonstructural Protein 4B on the Wnt/β-catenin pathway
BMC, 2017Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Jing RenAbstract:Abstract Background Hepatitis C virus (HCV) core Protein and Nonstructural Protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core Protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. Results Compared with the empty vector control, HCV core Protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P
-
screening of differentially expressed genes and gene pathways in hepatitis c virus 1b type Nonstructural Protein 4B stably expressed l02 cell line
Journal of Central South University. Medical sciences, 2014Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Chuang Lei, Bo Jiang, Hua PengAbstract:OBJECTIVE To screen differentially expressed genes and gene pathways in L02 cell line stably expressing hepatitis C virus (HCV) Ib type Nonstructural Protein 4B (NS4B) mediated by lentiviral system, and to provide a basis for further research of molecular biological mechanism of NS4B gene in chronic hepatitis C and hepatocarcinogenesis. METHODS NS4B stably overexpressed L02 cell line and negative control stable L02 cell line, designated as L02-NS4B and L02-mkate2 respectively, were resurrected and amplified in vitro. The differentially expressed genes between L02-NS4B and L02-mkate2 were determined by gene expression microarray from Human Gene 1.0ST. The significant pathways of the differential genes were selected by the Fisher's exact test and χ2 test according to kyoto encyclopedia of genes and genomes (KEGG) database. The differential expression levels of 5 selected genes including Protein kinase C delta binding Protein (PRKCDBP), tumor Protein p53 (TP53), v-akt murine thymoma viral oncogene homolog 1 (AKT1), baculoviral IAP repeat containing 3 (BIRC3) and B-cell lymphoma 2-like1 (BCL2L1) from cDNA microarray data were further verified by real-time quantitative polymerase chain reaction (real-time QPCR). RESULTS Between L02-NS4B and L02-mkate2, the genes with flurescence intensity ratio >1.2 or <0.8 were considered as differentially expressed genes. A total of 2 682 differentially expressed genes in the known 28 869 human genes were detected in L02-NS4B, 1 446 genes were upregulated and 1 236 genes were downregulated. A total of 41 involved pathways of up-regulated differential genes were identified by KEGG database, mainly including apoptosis, extracellular matrix receptor interaction, cell cycle, pathways in cancer and Toll-like receptor signaling pathway; and 20 involved pathways of down-regulated differential genes were identified, mainly including pathways in cancer, Wnt signaling pathway and cell cycle pathway. Of the 5 upregulated genes selected from cDNA microarray data, 3 genes showed the same differential expression pattern by real-time QPCR as that shown in cDNA microarray data, namely AKT1, BIRC3 and BCL2L1. The confirmation rate of real-time QPCR was 60%. CONCLUSION HCVNS4B can up-regulate or down-regulate the expression of many genes in L02 cells, thus affecting multiple signaling pathways relevant to cell apoptosis, cell cycle and carcinogenesis.
-
Screening of differentially expressed genes and gene pathways in hepatitis C virus 1b type Nonstructural Protein 4B stably expressed L02 cell line
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences, 2014Co-Authors: Xiaohua Jiang, Yu-tao Xie, Ya-ping Cai, Chuang Lei, Bo Jiang, Hua PengAbstract:OBJECTIVE To screen differentially expressed genes and gene pathways in L02 cell line stably expressing hepatitis C virus (HCV) Ib type Nonstructural Protein 4B (NS4B) mediated by lentiviral system, and to provide a basis for further research of molecular biological mechanism of NS4B gene in chronic hepatitis C and hepatocarcinogenesis. METHODS NS4B stably overexpressed L02 cell line and negative control stable L02 cell line, designated as L02-NS4B and L02-mkate2 respectively, were resurrected and amplified in vitro. The differentially expressed genes between L02-NS4B and L02-mkate2 were determined by gene expression microarray from Human Gene 1.0ST. The significant pathways of the differential genes were selected by the Fisher's exact test and χ2 test according to kyoto encyclopedia of genes and genomes (KEGG) database. The differential expression levels of 5 selected genes including Protein kinase C delta binding Protein (PRKCDBP), tumor Protein p53 (TP53), v-akt murine thymoma viral oncogene homolog 1 (AKT1), baculoviral IAP repeat containing 3 (BIRC3) and B-cell lymphoma 2-like1 (BCL2L1) from cDNA microarray data were further verified by real-time quantitative polymerase chain reaction (real-time QPCR). RESULTS Between L02-NS4B and L02-mkate2, the genes with flurescence intensity ratio >1.2 or
