Jalili Shirin, Hashemi Seyed Mohammad Ali, Sarvari Jamal
Institute of police equipment and technologies, policing sciences and social studies research institute, Tehran, Iran.
Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Rep Biochem Mol Biol. 2024 Apr;13(1):51-58. doi: 10.61186/rbmb.13.1.51.
Individuals experiencing severe cases of Coronavirus Disease 2019 (COVID-19) exhibited elevated fibrinogen levels and decreased albumin levels, potentially linked to the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) proteins. Consequently, our study endeavors to examine the impact of SARS-CoV-2 ORF9b on the expression of fibrinogen and albumin genes within the Hep-G2 cell line.
In this study, the Hep-G2 liver cell line was utilized alongside the plasmid pcDNA3.1 hyg+ containing ORF9b from the SARS-CoV-2 strain originating in Wuhan. Transfection procedures were executed, and the transfected cells were selected utilizing hygromycin B. Validation of ORF9b expression was conducted through SYBR green-based real-time PCR, and the expression of the Fibrinogen α (FGA), Fibrinogen β (FGB), Fibrinogen γ (FGG), and Albumin (ALB) genes was quantified using the same method.
The real-time PCR analysis revealed a significant upregulation of fibrinogen genes-α (P=0.03), β (P=0.02), and γ (P=0.029) in Hep-G2 cells containing ORF9b compared to control cells. Furthermore, the findings indicated a markedly lower expression level of albumin in Hep-G2 cells harboring ORF9b compared to the control cells (P=0.028).
The findings suggest that SARS-CoV-2 ORF9b could potentially influence the course of SARS-CoV-2 infection by triggering the expression of α, β, and γ fibrinogen gene chains while suppressing the albumin gene. Further investigations are warranted to validate these observations across various SARS-CoV-2 strains exhibiting differing levels of pathogenicity.
2019年冠状病毒病（COVID-19）重症患者的纤维蛋白原水平升高而白蛋白水平降低，这可能与严重急性呼吸综合征冠状病毒2（SARS-CoV-2）蛋白的存在有关。因此，我们的研究旨在探讨SARS-CoV-2 ORF9b对Hep-G2细胞系中纤维蛋白原和白蛋白基因表达的影响。
在本研究中，使用了Hep-G2肝细胞系以及含有源自武汉的SARS-CoV-2毒株ORF9b的质粒pcDNA3.1 hyg+。进行了转染程序，并使用潮霉素B筛选转染细胞。通过基于SYBR green的实时PCR验证ORF9b的表达，并使用相同方法定量纤维蛋白原α（FGA）、纤维蛋白原β（FGB）、纤维蛋白原γ（FGG）和白蛋白（ALB）基因的表达。
实时PCR分析显示，与对照细胞相比，含有ORF9b的Hep-G2细胞中纤维蛋白原基因α（P=0.03）、β（P=0.02）和γ（P=0.029）显著上调。此外，研究结果表明，与对照细胞相比，含有ORF9b的Hep-G2细胞中白蛋白的表达水平明显较低（P=0.028）。
研究结果表明，SARS-CoV-2 ORF9b可能通过触发α、β和γ纤维蛋白原基因链表达同时抑制白蛋白基因来影响SARS-CoV-2感染进程。有必要进行进一步研究以在具有不同致病性水平的各种SARS-CoV-2毒株中验证这些观察结果。
