Chumchanchira Chanida, Sornjai Wannapa, Roytrakul Sittiruk, Lithanatudom Pathrapol, Smith Duncan R
PhD Degree Program in Biology, Faculty of Science, Chiang Mai University, Chiang Mai, Thailand.
Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, Thailand.
PLoS One. 2025 Aug 1;20(8):e0329783. doi: 10.1371/journal.pone.0329783. eCollection 2025.
The mosquito transmitted dengue virus (DENV; family Flaviviridae, genus Orthoflavivirus, species Orthoflavivirus denguei) is a significant public health problem in many tropical and subtropical countries around the world. Human infection by DENV is predominantly asymptomatic in 80% of cases, but the remaining 20% of infections can result in symptoms ranging from a mild undifferentiated fever to life threatening dengue hemorrhagic and dengue shock syndrome. During infection DENV induces changes in the host cell, including changing protein expression, altering the cellular lipids and inducing changes in membrane architecture. A number of cell types have been shown to be permissive for DENV replication, including hepatocytes. This study sought to investigate the protein expression changes induced by DENV infection of a liver cell line, Huh-7, using 2-dimensional (2D) electrophoresis. At 48 hours post infection 14 protein spots were found to have altered expression as compared to mock infected cells at the same time point. In particular four of the proteins showing alterations of expression were chaperone proteins (Stress-70 protein, Endoplasmic reticulum chaperone BiP (GRP78), Heat shock 70 kDa protein 4 and Heat shock protein HSP 90-beta), of which three were upregulated (Stress-70 protein, Endoplasmic reticulum chaperone BiP (GRP78), Heat shock 70 kDa protein 4) and one was downregulated (Heat shock protein HSP 90-beta). GRP78 showed the largest change in expression amongst these four proteins, and so its expression was confirmed by western blot analysis. GRP78 has been shown by many studies to be critically involved in the replication of orthoflaviviruses, and this study further underlines the importance of this protein.
蚊子传播的登革病毒（DENV；黄病毒科，正黄病毒属，登革正黄病毒种）是世界上许多热带和亚热带国家的一个重大公共卫生问题。在80%的病例中，人类感染DENV主要表现为无症状，但其余20%的感染可导致从轻度未分化发热到危及生命的登革出血热和登革休克综合征等症状。在感染过程中，DENV会诱导宿主细胞发生变化，包括改变蛋白质表达、改变细胞脂质以及诱导膜结构变化。已证明多种细胞类型可允许DENV复制，包括肝细胞。本研究旨在利用二维（2D）电泳研究DENV感染肝细胞系Huh-7诱导的蛋白质表达变化。感染后48小时，与同一时间点的模拟感染细胞相比，发现14个蛋白点的表达发生了改变。特别值得注意的是，表达发生改变的四种蛋白质是伴侣蛋白（应激70蛋白、内质网伴侣BiP（GRP78）、热休克70 kDa蛋白4和热休克蛋白HSP 90-β），其中三种上调（应激70蛋白、内质网伴侣BiP（GRP78）、热休克70 kDa蛋白4），一种下调（热休克蛋白HSP 90-β）。GRP78在这四种蛋白质中表达变化最大，因此通过蛋白质印迹分析证实了其表达。许多研究表明GRP78在正黄病毒的复制中起关键作用，本研究进一步强调了该蛋白的重要性。
