Table_1_Overexpression of GINS4 Is Associated With Tumor Progression and Poor Survival in Hepatocellular Carcinoma.docx

PurposeOur research was aimed to identify the expression, clinical value and biological significance of GINS complex subunit 4 (GINS4) in hepatocellular carcinoma (HCC).
Materials and MethodsGINS4 was initially screened through weighted gene co-expression network analysis (WGCNA). The TCGA, GEO, and TIMER databases were applied for analyzing the GINS4 mRNA expression in HCC. GINS4 protein levels were detected via immunohistochemistry (IHC). Receiver operating characteristic (ROC) curve was applied for estimating the diagnostic significance of GINS4 in HCC. Kaplan-Meier plots, Cox model, and nomogram were used to assess the prognostic performance of GINS4 in HCC. Nomogram validation was conducted through time-dependent ROC and decision curve analysis (DCA). The Wanderer, UALCAN, and DiseaseMeth databases were utilized to identify GINS4 methylation levels in HCC. Genes co-expressed with GINS4 in HCC were estimated through the TCGA, cBioPortal, and GEPIA. GO, KEGG, and GSEA unraveled the possible biological mechanisms of GINS4 in HCC.
ResultsWGCNA confirmed that GINS4 was one of hub genes significantly associated with histological grade of HCC. Multiple databases confirmed the significant upregulation of GINS4 in HCC tissues compared with non-tumor controls. IHC analysis of 35 HCC patients demonstrated that overexpressed GINS4 positively correlated with advanced TNM stage and poor pathological differentiation. GINS4 could effectively differentiate HCC cases from healthy individuals, with an AUC of 0.865. Increased GINS4 expression predicted unsatisfactory prognosis in HCC patients, especially in age >60 years, histological grade 1, HBV infection-negative, and occurring relapse subgroup. Nomogram incorporating GINS4 level and TNM stage displayed satisfactory predictive accuracy and clinical utility in predicting HCC prognosis. Upregulated GINS4 exhibited hypomethylated levels in HCC. Functional analysis indicated that GINS4 potentially positively modulated cell cycle and PI3K/AKT/mTOR pathway.
ConclusionGINS4 is overexpressed in HCC and is correlated with undesirable survival of HCC patients.

研究目的：本研究旨在明确GINS复合物亚基4（GINS complex subunit 4, GINS4）在肝细胞癌（hepatocellular carcinoma, HCC）中的表达情况、临床价值及生物学意义。
材料与方法：本研究首先通过加权基因共表达网络分析（weighted gene co-expression network analysis, WGCNA）筛选GINS4。采用TCGA、GEO及TIMER数据库分析肝细胞癌中GINS4的mRNA表达水平。通过免疫组化（immunohistochemistry, IHC）检测GINS4的蛋白表达水平。采用受试者工作特征（receiver operating characteristic, ROC）曲线评估GINS4在肝细胞癌中的诊断价值。采用Kaplan-Meier绘图法、Cox模型及列线图（nomogram）评估GINS4在肝细胞癌中的预后效能。通过时间依赖性ROC曲线及决策曲线分析（decision curve analysis, DCA）对列线图进行验证。采用Wanderer、UALCAN及DiseaseMeth数据库分析肝细胞癌中GINS4的甲基化水平。通过TCGA、cBioPortal及GEPIA数据库筛选与肝细胞癌中GINS4共表达的基因。通过GO富集分析、KEGG富集分析及GSEA富集分析阐明GINS4在肝细胞癌中潜在的生物学机制。
研究结果：加权基因共表达网络分析证实GINS4为与肝细胞癌组织学分级显著相关的核心基因之一。多数据库分析证实，相较于非肿瘤对照组织，肝细胞癌组织中GINS4的表达显著上调。对35例肝细胞癌患者的免疫组化分析显示，GINS4高表达与晚期TNM分期及较差的病理分化程度呈正相关。GINS4可有效区分肝细胞癌患者与健康个体，曲线下面积（area under curve, AUC）达0.865。GINS4高表达预示肝细胞癌患者预后不良，尤其在年龄＞60岁、组织学分级1级、HBV感染阴性及复发亚组患者中更为显著。整合GINS4表达水平与TNM分期的列线图在预测肝细胞癌预后方面展现出良好的预测精度与临床实用性。肝细胞癌中GINS4高表达伴随低甲基化水平。功能分析表明，GINS4可能正向调控细胞周期及PI3K/AKT/mTOR信号通路。
研究结论：GINS4在肝细胞癌中呈高表达状态，且与肝细胞癌患者的不良生存结局显著相关。