基于生物信息学分析PNCK在肝癌中的表达及其与预后、免疫浸润的相关性

To analyze the expression of PNCK in hepatocellular carcinoma and its correlation with prognosis and immune infiltration based on bioinformatics

目的:采用生物信息学方法探讨妊娠上调的非泛素性钙调蛋白激酶(PNCK)在肝癌中的表达及其与预后、免疫浸润的关系。方法:基于TCGA数据库分析PNCK mRNA在肝癌中的表达,采用Cox回归分析肝癌患者预后的危险因素,GEPIA2数据库分析PNCK mRNA表达水平与肝癌患者中位生存时间的相关性,HPA数据库分析PNCK蛋白在肝癌组织及正常肝脏组织中的表达情况,TIMER数据库分析PNCK与肝癌组织中免疫细胞浸润程度的相关性,LinkedOmics数据库鉴定出肝癌组织中与PNCK的共表达基因,共表达基因的GO、KEGG富集分析采用DAVID数据库分析。结果:肝癌组织中PNCK mRNA表达水平为0.041(0.010,0.365),正常组织中PNCK mRNA表达水平为0(0,0.015)(P<0.001);50例肝癌组织及配对的癌旁组织,肝癌组织中PNCK mRNA表达水平为0.028(0,0.194),正常组织中PNCK mRNA表达水平为0(0,0.015)(P<0.001)。Cox多因素分析结果显示,PNCK mRNA表达水平(高表达)是影响肝癌患者预后的危险因素(HR=1.827,95%CI=1.145~2.914,P=0.011),K-M曲线结果证实,PNCK mRNA高表达的肝癌患者中位生存时间显著降低(P=0.002)。PNCK蛋白在肝癌组织中呈高表达,在正常肝脏组织中呈低表达,主要表达于细胞质中。肝癌组织中PNCK mRNA表达水平与Th2细胞、TFH细胞、NK CD56bright细胞等呈显著正相关(P<0.001),与DC细胞、Neutrophils、Th17细胞等呈显著负相关(P<0.001)。肝癌组织中PNCK共表达基因主要富集于胰岛素分泌调节、线粒体基质、氧化还原活动、代谢途径、丙酮酸代谢、糖酵解和糖异生等。结论:PNCK在肝癌中显著高表达,可作为肝癌预后不良的标志物,PNCK可能通过影响免疫细胞浸润及代谢途径调控肝癌的发生发展。

Objective:To investigate the expression of PNCK in hepatocellular carcinoma(HCC)and its relationship with prognosis,immune infiltration by bioinformatics method.Methods:The expression of PNCK mRNA in HCC was analyzed based on TCGA database,and the prognostic risk factors of HCC patients were analyzed by Cox regression.GEPIA2 database was used to analyze the correlation between PNCK mRNA expression level and median survival time of patients with HCC.The expression of PNCK protein in HCC tissues and normal liver tissues was analyzed by HPA database.TIMER database was used to analyze the correlation between PNCK and the degree of immune cell infiltration in HCC tissues.The co-expressed genes with PNCK in HCC tissues was identified by LinkedOmics database.The GO and KEGG enrichment analysis of co-expressed genes was performed using DAVID database.Results:The expression level of PNCK mRNA in HCC tissues and normal liver tissues was 0.041(0.010,0.365)and 0.000(0.000,0.015),respectively.Compared with normal liver tissues,the expression level of PNCK mRNA in HCC tissues was significantly higher(P<0.001).The PNCK mRNA expression level in HCC tissues and paracancerous tissues was 0.028(0.000,0.194)and 0.000(0.000,0.015),respectively.The mRNA expression level of PNCK in HCC tissues was also significantly higher(P<0.001).The Cox multivariate analysis showed that PNCK mRNA expression level(high expression)was a risk factor affecting the prognosis of patients with HCC(HR=1.827,95%CI:1.145-2.914,P=0.011).K-M curve results confirmed that the median survival time of HCC patients with high expression of PNCK mRNA was significantly reduced(P=0.002).The PNCK protein was highly expressed in HCC tissues and low expressed in normal liver tissues,which mainly expressed in cytoplasm.The mRNA expression level of PNCK in HCC tissues was positively correlated with Th2 cells,TFH cells and NK CD56bright cells(P<0.001),and negatively correlated with DC cells,Neutrophils and Th17 cells(P<0.001).PNCK co-expressed genes in HCC were mainly enriched in positive regulation of insulin secretion,mitochondrial matrix,oxidoreductase activity,metabolic pathways,pyruvate metabolism,glycolysis/gluconeogenesis.Conclusion:PNCK is over-expressed in HCC,and can be used as a marker of poor prognosis of HCC.The PNCK could regulate immune cell infiltration and metabolic pathways which may involved the pathogenesis of HCC.