基于生物信息学分析核仁纺锤体相关蛋白1在肝细胞癌中的表达及对临床预后的影响

Expression of nucleolar spindle-associated protein 1 in hepatocellular carcinoma and its effect on clinical prognosis:A bioinformatics analysis

目的探讨在肝癌中核仁纺锤体相关蛋白1(NUSAP1)的表达及其对临床预后的影响。方法从基因表达数据库(GEO)下载HCC芯片数据集GSE57957、GSE14520、GSE22058、GSE46444、GSE54236、GSE36376、GSE64041、GSE76297、GSE76427、GSE102079和癌症基因组图谱(TCGA)中的HCC RNA-seq数据,利用R 4.0软件分析NUSAP1在肝癌及癌旁组织的表达差异。应用t-分布邻域嵌入算法(tSNE)对单细胞测序数据的降维,分析NUSAP1在肝癌和癌旁组织中的差异表达。收集2018年1月—2019年11月在青岛大学附属医院进行同种异体肝移植术的42例乙型肝炎相关肝癌患者的癌组织及癌旁组织,收集其临床病理资料并分析与NUSAP1基因表达的相关性。计数资料2组间比较采用χ^(2)检验。以Kaplan-Meier方法生存分析NUSAP1表达水平与总体生存率及无病间隔生存率之间的关系,运用log-rank进行检验,通过Graphpad prim 7使其结果可视化。结果NUSAP1在GEO数据库与TCGA数据库数据集中的肝癌组织中均高表达(P值均<0.001)。来源于肿瘤组织的细胞其NUSAP1高表达细胞明显高于癌旁组织细胞。NUSAP1在29例患者的肝癌组织中高表达,在13例患者的肝癌组织中低表达,NUSAP1在肝癌组织中高表达组及低表达组间,Child-Pugh分级、TNM分期、Okuda分期比较差异均有统计学意义(χ^(2)值分别为5.469、6.836、4.617,P值均<0.05)。NUSAP1高表达的肝癌患者总体生存率与无病间隔生存率显著低于低表达的肝癌患者,NUSAP1低表达患者1、3、5生存率分别为92.96%、83.80%、76.76%,明显高于高表达的患者(76.76%、67.60%、64.78%),NUSAP1高表达的患者中位生存时间明显低于低表达的患者(61.7个月vs 108.6个月)(P值均<0.05)。结论NUSAP1在肝癌细胞中高表达,可作为乙型肝炎肝癌患者不良预后的潜在标志物。

Objective To investigate the expression of the nucleolar spindle-associated protein 1(NUSAP1)gene in hepatocellular carcinoma(HCC)and its effect on clinical prognosis.Methods HCC microarray datasets GSE57957,GSE14520,GSE22058,GSE46444,GSE54236,GSE36376,GSE64041,GSE76297,GSE76427,and GSE102079 were downloaded from the Gene Expression Omnibus(GEO)database,and HCC RNA-Seq data were downloaded from The Cancer Genome Atlas(TCGA).R software 4.0 was used to investigate the difference in the expression of NUSAP1 between HCC tissue and adjacent tissue.The t-distributed stochastic neighbour embedding(t-SNE)method was used to perform dimensionality reduction of single-cell sequencing data and analyze the difference in the expression of NUSAP1 between HCC tissue and adjacent tissue.HCC tissue samples and adjacent tissue samples were collected from 42 patients with hepatitis B-related HCC who underwent liver transplantation in The Affiliated Hospital of Qingdao University from January 2018 to November 2019,and related clinical and pathological data were collected to analyze their correlation with the expression of the NUSAP1 gene.The chi-square test was used for comparison of paired data.The Kaplan-Meier method was used to analyze the correlation of the expression level of NUSAP1 with overall survival rate and disease-free survival,and the log-rank test was used for survival difference analysis;the results were visualized using Graphpad prim 7.Results NUSAP1 was highly expressed in HCC tissue in GEO and TCGA datasets(P<0.001).The cells from HCC tissue had a significantly higher proportion of cells with high NUSAP1 expression than those from adjacent tissue.NUSAP1 showed high expression in HCC tissue of 29 patients and low expression in HCC tissue of 13 patients,and there were significant differences in Child-Pugh class,TNM stage,and Okuda stage between the high-expression group and the low-expression group(χ^(2)=5.469,6.836 and 4.617,all P<0.05).The HCC patients with high expression of NUSAP1 had significantly lower overall survival rate and disease-free survival rate than those with low expression;compared with the patients with high expression,the patients with low expression had significantly higher 1-,3-,and 5-year survival rates(92.96%/83.80%/76.76%vs 76.76%/67.60%/64.78%,P<0.05),and the patients with high expression had a significantly shorter median survival time than those with low expression(61.7 months vs 108.6 months,P<0.05).Conclusion NUSAP1 is highly expressed in cells from HCC tissue and may be used as a potential marker for poor prognosis of patients with hepatitis B-related HCC.