关键词: Chronic hepatitis C virus (HCV) infection Direct-acting antiviral agents (DAA) Endoplasmic reticulum stress (ER stress) Glycogen synthase kinase-3β (GSK-3β) Hepatocellular carcinoma (HCC) Protein kinase A (PKA) Wnt/β-catenin

Mesh : Humans Antiviral Agents / pharmacology beta Catenin Carcinoma, Hepatocellular Glycogen Synthase Kinase 3 beta Hepacivirus Hepatitis C, Chronic / drug therapy Liver Neoplasms Endoplasmic Reticulum Stress Protein Kinase Inhibitors / pharmacology Cells, Cultured

来  源:   DOI:10.1186/s12964-023-01081-9   PDF(Pubmed)

Abstract:
Chronic hepatitis C virus (HCV) infection causes hepatocellular carcinoma (HCC). The HCC risk, while decreased compared with active HCV infection, persists in HCV-cured patients by direct-acting antiviral agents (DAA). We previously demonstrated that Wnt/β-catenin signaling remained activated after DAA-mediated HCV eradication. Developing therapeutic strategies to both eradicate HCV and reverse Wnt/β-catenin signaling is needed.
Cell-based HCV long term infection was established. Chronically HCV infected cells were treated with DAA, protein kinase A (PKA) inhibitor H89 and endoplasmic reticulum (ER) stress inhibitor tauroursodeoxycholic acid (TUDCA). Western blotting analysis and fluorescence microscopy were performed to determine HCV levels and component levels involved in ER stress/PKA/glycogen synthase kinase-3β (GSK-3β)/β-catenin pathway. Meanwhile, the effects of H89 and TUDCA were determined on HCV infection.
Both chronic HCV infection and replicon-induced Wnt/β-catenin signaling remained activated after HCV and replicon eradication by DAA. HCV infection activated PKA activity and PKA/GSK-3β-mediated Wnt/β-catenin signaling. Inhibition of PKA with H89 both repressed HCV and replicon replication and reversed PKA/GSK-3β-mediated Wnt/β-catenin signaling in both chronic HCV infection and replicon. Both chronic HCV infection and replicon induced ER stress. Inhibition of ER stress with TUDCA both repressed HCV and replicon replication and reversed ER stress/PKA/GSK-3β-dependent Wnt/β-catenin signaling. Inhibition of either PKA or ER stress both inhibited extracellular HCV infection.
Targeting ER stress/PKA/GSK-3β-dependent Wnt/β-catenin signaling with PKA inhibitor could be a novel therapeutic strategy for HCV-infected patients to overcomes the issue of remaining activated Wnt/β-catenin signaling by DAA treatment. Video Abstract.
摘要:
背景:慢性丙型肝炎病毒(HCV)感染导致肝细胞癌(HCC)。HCC风险,虽然与活动性HCV感染相比有所下降,通过直接作用的抗病毒药物(DAA)在HCV治愈的患者中持续存在。我们先前证明,在DAA介导的HCV根除后,Wnt/β-catenin信号传导仍然被激活。需要开发根除HCV和逆转Wnt/β-catenin信号传导的治疗策略。
方法:建立基于细胞的HCV长期感染。慢性HCV感染的细胞用DAA处理,蛋白激酶A(PKA)抑制剂H89和内质网(ER)应激抑制剂牛磺熊去氧胆酸(TUDCA)。进行蛋白质印迹分析和荧光显微镜检查以确定涉及ER应激/PKA/糖原合酶激酶-3β(GSK-3β)/β-catenin途径的HCV水平和成分水平。同时,确定了H89和TUDCA对HCV感染的影响.
结果:慢性HCV感染和复制子诱导的Wnt/β-catenin信号在通过DAA根除HCV和复制子后保持激活。HCV感染激活PKA活性和PKA/GSK-3β介导的Wnt/β-catenin信号传导。在慢性HCV感染和复制子中,用H89抑制PKA既抑制HCV和复制子复制,又逆转PKA/GSK-3β介导的Wnt/β-连环蛋白信号传导。慢性HCV感染和复制子均诱导ER应激。用TUDCA抑制ER应激既抑制HCV和复制子复制,又逆转ER应激/PKA/GSK-3β依赖性Wnt/β-连环蛋白信号传导。抑制PKA或ER应激均抑制细胞外HCV感染。
结论:用PKA抑制剂靶向ER应激/PKA/GSK-3β依赖性Wnt/β-catenin信号传导可能是HCV感染患者的一种新的治疗策略,以克服DAA治疗仍然激活Wnt/β-catenin信号传导的问题。视频摘要。
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