The Paradoxical Effects of Different Hepatitis C Viral Loads on Host DNA Damage and Repair Abilities.
The Paradoxical Effects of Different Hepatitis C Viral Loads on Host DNA Damage and Repair Abilities.
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DOI：
10.1371/journal.pone.0164281
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Yu ML
中科院分区：
文献类型：
--
作者：
Wang SC;Lai KR;Li CY;Chiang CS;Yu GY;Sakamoto N;Tu WY;Hsieh MH;Huang JF;Chuang WL;Dai CY;Yu ML
丙型肝炎病毒（HCV）诱导的肝脏应激与DNA氧化损伤增加有关，并与肝脏炎症有关。然而，HCV感染和复制是不均匀的，并且在个体肝细胞中有所不同。为了研究病毒载量对宿主DNA损伤的影响，我们使用增强黄色荧光蛋白基因（EYFP）标记的HCV病毒来区分HCV细胞内高病毒载量（HVL）细胞和低病毒载量（LVL）细胞。HCV多蛋白的高表达证实了细胞分选效率。我们在HVL人群中发现了DNA损伤γ-H2AX灶。彗星试验表明，HVL与DNA链断裂的程度有关。令人惊讶的是，DNA qPCR阵列和western blotting显示，损伤相关基因GPX2、MRE11、phospho-ATM和OGG1在LVL细胞中显著上调，而在HVL细胞中呈负向下调或一致表达。检测这些细胞在辐照下修复能力的菌落存活试验表明，LVL细胞对辐照的抵抗力更强，修复辐射引起的损伤的能力也更强。该研究发现，细胞内病毒载量驱动细胞DNA损伤水平，但抑制损伤相关基因表达。然而，LVL细胞中损伤相关基因表达的增加可能受到HVL细胞ROS的影响。这些发现为hcv感染细胞中不同病毒载量导致的不同DNA损伤和修复反应提供了新的见解。
Hepatitis C virus (HCV)-induced hepatic stress is associated with increased oxidative DNA damage and has been implicated in hepatic inflammation. However, HCV infection and replication are uneven and vary among individual hepatocytes. To investigate the effect of the viral load on host DNA damage, we used an Enhanced Yellow Fluorescent Protein gene (EYFP)-tagged HCV virus to distinguish between HCV intracellular high viral load (HVL) cells and low viral load (LVL) cells. The cell sorting efficiency was confirmed by the high expression of the HCV polyprotein. We found DNA damage γ-H2AX foci in the HVL population. Comet assays demonstrated that HVL was related to the extent of the DNA strand breaks. Surprisingly, the DNA qPCR arrays and western blotting showed that the damage-related genes GPX2, MRE11, phospho-ATM, and OGG1 were significantly up-regulated in LVL cells but inversely down-regulated or consistently expressed in HVL cells. The colony survival assay to examine the repair abilities of these cells in response to irradiation showed that the LVL cells were more resistant to irradiation and had an increased ability to repair radiation-induced damage. This study found that intracellular viral loads drove cellular DNA damage levels but suppressed damage-related gene expression. However, the increase in damage-related gene expression in the LVL cells may be affected by ROS from the HVL cells. These findings provide new insights into the distinct DNA damage and repair responses resulting from different viral loads in HCV-infected cells.
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影响因子：
9.7
作者：
Chen, Benjamin P. C.;Li, Mengxia;Asaithamby, Aroumougame
通讯作者：
Asaithamby, Aroumougame
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作者：
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