Zhu Wenjie, Xu Jian, Chen Zehao, Jiang Jianxin
Department of Hepatobiliary Surgery, Renmin Hospital of Wuhan University, Wuhan, China.
Front Genet. 2021 Jul 20;12:689159. doi: 10.3389/fgene.2021.689159. eCollection 2021.
Hepatocellular carcinoma (HCC) is one of the most common carcinomas worldwide. Our study aims to analyze how NUSAP1 affects progression of HCC from clinical, molecular mechanism and immune perspectives. Firstly, we downloaded GSE62232, GSE102079, GSE112790, and GSE121248 gene expression profile datasets from GEO database. R studio was used to screen DEGs of each dataset, and 86 overlapping DEGs of the four datasets were screened at last. Then, CytoHubba plug-in in Cytoscape software was used to screen out NUSAP1 from the 86 DEGs. Subsequently, survival analysis, clinical correlation analysis, independent prognostic analysis, and GSEA enrichment analysis of NUSAP1 were analyzed using HCC patients from GSE76427 dataset, ICGC database, and TCGA database. The results revealed that HCC patients with higher expression level of NUSAP1 had a worse prognosis. NUSAP1 was an independent prognostic factor of HCC, and it may promote HCC progress by regulating cell cycle. To further elucidate its underlying molecular mechanism, we used cBioProtal online data analysis tool to screen all co-expression genes of NUSAP1 and used top 300 co-expression genes to accomplish KEGG and GO enrichment analysis; the results confirmed that NUSAP1 accelerated progression of HCC by regulating cell cycle. We continued to draw KEGG pathway map of cell cycle using co-expression genes enriched in cell cycle pathway by KEGG online tool. The map depicted that most of co-expression genes of NUSAP1 were located in S phase and G2/M phase of the cell cycle, and they could regulate the genes in G1 phase. To further understand the mechanism of cell cycle, we also did qRT-PCR, Western blot, and flow cytometry; the results showed that NUSAP1 was closely associated with CDK4, CDK6, and cyclinD1, which could regulate G1 to S phase transition. Besides, we also analyzed correlation between NUSAP1 and immune cells using HCC patients from GSE76427 dataset, ICGC database, and TCGA database. NUSAP1 was associated with some immune cells, and we speculated that NUSAP1 could also promote HCC progression by influencing T cell CD4 memory resting and macrophage M0 through some underlying mechanism.
肝细胞癌（HCC）是全球最常见的癌症之一。我们的研究旨在从临床、分子机制和免疫角度分析NUSAP1如何影响HCC的进展。首先，我们从GEO数据库下载了GSE62232、GSE102079、GSE112790和GSE121248基因表达谱数据集。使用R studio筛选每个数据集的差异表达基因（DEG），最终筛选出四个数据集的86个重叠DEG。然后，使用Cytoscape软件中的CytoHubba插件从86个DEG中筛选出NUSAP1。随后，使用来自GSE76427数据集、ICGC数据库和TCGA数据库的HCC患者对NUSAP1进行生存分析、临床相关性分析、独立预后分析和GSEA富集分析。结果显示，NUSAP1表达水平较高的HCC患者预后较差。NUSAP1是HCC的独立预后因素，它可能通过调节细胞周期促进HCC进展。为了进一步阐明其潜在的分子机制，我们使用cBioProtal在线数据分析工具筛选NUSAP1的所有共表达基因，并使用前300个共表达基因完成KEGG和GO富集分析；结果证实NUSAP1通过调节细胞周期加速HCC进展。我们继续使用KEGG在线工具，利用富集在细胞周期途径中的共表达基因绘制细胞周期的KEGG途径图。该图显示，NUSAP1的大多数共表达基因位于细胞周期的S期和G2/M期，它们可以调节G1期的基因。为了进一步了解细胞周期机制，我们还进行了qRT-PCR、蛋白质免疫印迹和流式细胞术；结果表明，NUSAP1与CDK4、CDK6和细胞周期蛋白D1密切相关，它们可以调节G1期到S期的转变。此外，我们还使用来自GSE76427数据集、ICGC数据库和TCGA数据库的HCC患者分析了NUSAP1与免疫细胞之间的相关性。NUSAP1与一些免疫细胞有关，我们推测NUSAP1也可能通过某种潜在机制影响T细胞CD4记忆静息细胞和巨噬细胞M0，从而促进HCC进展。
