RNA-seq based transcriptome analysis of hepatitis E virus (HEV) and hepatitis B virus (HBV) replicon transfected Huh-7 cells.
RNA-seq based transcriptome analysis of hepatitis E virus (HEV) and hepatitis B virus (HBV) replicon transfected Huh-7 cells.
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基于RNA-SEQ的转录组分析乙型肝炎病毒（HEV）和丙型肝炎病毒（HBV）复制子转染了HUH-7细胞。
DOI：
10.1371/journal.pone.0087835
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Panda SK
中科院分区：
文献类型：
--
作者：
Jagya N;Varma SP;Thakral D;Joshi P;Durgapal H;Panda SK
乙型肝炎病毒(乙肝病毒)和戊型肝炎病毒(HEV)感染的发病机制千差万别，尽管它们看起来很相似；而乙肝病毒会导致急性和/或慢性肝病和肝细胞癌，而HEV主要引起急性自限性疾病。在这两种感染中，宿主反应在疾病建立和/或病毒清除中至关重要。随着HEV重叠感染对慢性乙肝预后的恶化，我们研究了仅HEV复制子(仅HEV)、仅HBV复制子(仅HBV单复制)以及HBVHEV和HBVHEV复制子(HBVHEV+HEV)对肝细胞(HuH-7细胞系)基因表达的影响。病毒复制采用链特异性实时RT-PCR检测HEV，培养上清液检测HBVHBs。间接免疫荧光对各自的病毒蛋白确认感染。通过RNA测序(RNA-Seq)分析转基因细胞中富含Poly-A的RNA，进行转录图谱分析。对每个样本中560万个碱基的平均读数进行了测序，并通过至少一次或多次读数绘制了∼15,800个基因的图谱。与模拟转染组相比，共有461个基因在HBVHEV组、408个HBV组和306个HEV组有差异表达(p<；0.05)或更多。大多数表达改变的重要基因聚集在免疫相关、信号转导和代谢过程类别中。基于实时定量RT-PCR的相关基因表达分析也验证了这些类别中功能重要基因的差异基因表达。据我们所知，这是首次报道体外复制子转染的RNA-Seq转录组分析，以了解宿主对HEV和乙肝病毒的反应。
Pathogenesis of hepatitis B virus (HBV) and hepatitis E virus (HEV) infection is as varied as they appear similar; while HBV causes an acute and/or chronic liver disease and hepatocellular carcinoma, HEV mostly causes an acute self-limiting disease. In both infections, host responses are crucial in disease establishment and/or virus clearance. In the wake of worsening prognosis described during HEV super-infection over chronic HBV hepatitis, we investigated the host responses by studying alterations in gene expression in liver cells (Huh-7 cell line) by transfection with HEV replicon only (HEV-only), HBV replicon only (HBV-only) and both HBV and HEV replicons (HBV+HEV). Virus replication was validated by strand-specific real-time RT-PCR for HEV and HBsAg ELISA of the culture supernatants for HBV. Indirect immunofluorescence for the respective viral proteins confirmed infection. Transcription profiling was carried out by RNA Sequencing (RNA-Seq) analysis of the poly-A enriched RNA from the transfected cells. Averages of 600 million bases within 5.6 million reads were sequenced in each sample and ∼15,800 genes were mapped with at least one or more reads. A total of 461 genes in HBV+HEV, 408 in HBV-only and 306 in HEV-only groups were differentially expressed as compared to mock transfection control by two folds (p<0.05) or more. Majority of the significant genes with altered expression clustered into immune-associated, signal transduction, and metabolic process categories. Differential gene expression of functionally important genes in these categories was also validated by real-time RT-PCR based relative gene-expression analysis. To our knowledge, this is the first report of in vitro replicon transfected RNA-Seq based transcriptome analysis to understand the host responses against HEV and HBV.
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影响因子：
3.7
作者：
Chandra V;Holla P;Ghosh D;Chakrabarti D;Padigaru M;Jameel S
通讯作者：
Jameel S
影响因子：
3.1
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Dorfleutner, Andrea;Bryan, Nicole B.;Stehlik, Christian
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Stehlik, Christian
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4.8
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Alnemri, ES
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Harton, Jonathan A.
影响因子：
2.5
作者：
Gupta, P.;Jagya, N.;Panda, S. K.
通讯作者：
Panda, S. K.