Steinkellner Hannes, Madritsch Silvia, Kluge Mara, Seipel Teresa, Sarne Victoria, Huber Anna, Schosserer Markus, Oberle Raimund, Neuhaus Winfried, Beribisky Alexander V, Laccone Franco
Center for Pathobiochemistry and Genetics, Institute of Medical Genetics, Medical University of Vienna, 1090 Vienna, Austria.
Vienna Doctoral School of Pharmaceutical, Nutritional and Sport Sciences (PhaNuSpo), University of Vienna, 1090 Vienna, Austria.
Genes (Basel). 2024 Nov 26;15(12):1514. doi: 10.3390/genes15121514.
BACKGROUND/OBJECTIVES: Nucleolin is a major component of the nucleolus and is involved in various aspects of ribosome biogenesis. However, it is also implicated in non-nucleolar functions such as cell cycle regulation and proliferation, linking it to various pathologic processes. The aim of this study was to use differential gene expression analysis and Weighted Gene Co-expression Network analysis (WGCNA) to identify nucleolin-related regulatory pathways and possible key genes as novel therapeutic targets for cancer, viral infections and other diseases.
We used two different siRNAs to downregulate the expression of nucleolin in a human hepatoblastoma (HepG2) cell line. We carried out RNA-sequencing (RNA-Seq), performed enrichment analysis of the pathways of the differentially expressed genes (DEGs) and identified protein-protein interaction (PPI) networks.
Both siRNAs showed high knockdown efficiency in HepG2 cells, resulting in the disruption of the nucleolar architecture and the downregulation of rRNA gene expression, both downstream hallmarks of a loss of nucleolin function. RNA-Seq identified 44 robust DEGs in both siRNA cell models. The enrichment analysis of the pathways of the downregulated genes confirmed the essential role of nucleolin in DNA replication and cell cycle processes. In addition, we identified seven hub genes linked to : , , , , , and ; all are known to be implicated in DNA replication, cell cycle progression and oncogenesis.
Our findings demonstrate the functional consequences of nucleolin depletion in HepG2 and confirm the importance of nucleolin in DNA replication and cell cycle processes. These data will further enhance our understanding of the molecular and pathologic mechanisms of nucleolin and provide new therapeutic perspectives in disease.
背景/目的：核仁素是核仁的主要成分，参与核糖体生物合成的各个方面。然而，它也涉及非核仁功能，如细胞周期调控和增殖，将其与各种病理过程联系起来。本研究的目的是使用差异基因表达分析和加权基因共表达网络分析（WGCNA）来识别与核仁素相关的调控途径和可能的关键基因，作为癌症、病毒感染和其他疾病的新型治疗靶点。
我们使用两种不同的小干扰RNA（siRNA）下调人肝癌细胞系（HepG2）中核仁素的表达。我们进行了RNA测序（RNA-Seq），对差异表达基因（DEG）的途径进行了富集分析，并确定了蛋白质-蛋白质相互作用（PPI）网络。
两种siRNA在HepG2细胞中均显示出高敲低效率，导致核仁结构破坏和rRNA基因表达下调，这两者都是核仁素功能丧失的下游标志。RNA-Seq在两种siRNA细胞模型中均鉴定出44个稳定的DEG。下调基因途径的富集分析证实了核仁素在DNA复制和细胞周期过程中的重要作用。此外，我们确定了七个与……、……、……、……、……、……和……相关的枢纽基因；已知所有这些基因都与DNA复制、细胞周期进程和肿瘤发生有关。
我们的研究结果证明了HepG2细胞中核仁素缺失的功能后果，并证实了核仁素在DNA复制和细胞周期过程中的重要性。这些数据将进一步加深我们对核仁素分子和病理机制的理解，并为疾病提供新的治疗视角。
