摘要
目的 基于GEPIA数据库探讨正常组织与肺腺癌组织的差异表达基因以及差异表达基因与肺腺癌分期的关系,Ka-plan-Meier Plotter数据库分析该基因对肺腺癌患者生存预后的影响和TIMER数据库分析差异表达基因与肿瘤免疫细胞浸润的关系.方法 通过使用GEPIA数据库来获得正常组织和肺腺癌的差异表达基因以及获取差异表达基因与肺腺癌分期的关系,通过使用Kaplan-Meier Plotter数据库对这些基因进行生存预后分析和通过使用TIMER数据库对差异表达基因进行肿瘤免疫细胞浸润分析;使用实时荧光定量PCR(qRT-PCR)验证FAM83A在肺腺癌细胞中的表达水平.结果 在GEPIA中,肺腺癌差异表达基因前 10 位分别为:RP11-40C6.2、XAGE1A、XAGE1B、RP5-940J5.9、IGHGP、CRABP2、FAM83A、SPP1、TM-PRSS4、CEACAM5,均在肺腺癌中呈高表达水平;其中IGHGP、FAM83A基因表达水平与肺腺癌分期相关,而RP11-40C6.2、XA-GE1A、XAGE1B、RP5-940J5.9、CRABP2、SPP1、TMPRSS4、CEACAM5 基因表达水平与肺腺癌分期无显著相关;在Kaplan-Meier Plotter生存分析中,XAGE1A、XAGE1B、CRABP2、FAM83A、SPP1、TMPRSS4 表达水平与 OS 呈负相关;XAGE1A、XAGE1B、CRABP2、FAM83A、SPP1 表达水平与 FP 呈负相关;FAM83A 表达水平与 PPS 呈负相关.在 TIMER 免疫细胞浸润分析中,CRABP2 与CD4+T细胞、巨噬细胞、中性粒细胞、树突状细胞呈正相关,与肿瘤纯度呈负相关;FAM83A与B细胞、树突状细胞呈负相关;SPP1 与巨噬细胞、中性粒细胞、树突状细胞呈正相关,与肿瘤纯度呈负相关;TMPRSS4 与CD4+T细胞呈正相关,与CD8+T细胞呈负相关;CEACAM5 与CD4+T细胞呈正相关,与CD8+T细胞呈负相关.qRT-PCR实验结果显示,FAM83A在肺腺癌细胞中呈高表达水平,差异具有统计学意义(P<0.05).结论 肺腺癌组织与正常组织之间存在差异表达基因,其中FAM83A在肺腺癌细胞中呈高表达水平,与肺腺癌患者分期、免疫细胞浸润、预后不良相关,可能是预测生存预后的一个潜在生物标志物.
Abstract
Objective To explore the differentially expressed genes between normal and lung adenocarcinoma(LUAD)tissues based on the GEPIA database and the relationship between differentially expressed genes and LUAD staging.The Kaplan Meier Plotter database was used to analyze this gene's impact on LUAD patients'survival prognosis,and the TIMER database was used to explore the relationship between differentially expressed genes and tumor immune cell infiltration.Methods In this study,we utilized the GE-PIA database to identify genes that are differentially expressed in normal lung tissue versus LUAD tissue.We also investigated the rela-tionship between these differentially expressed genes and the staging of LUAD.Next,we analyzed the prognostic value of these genes u-sing the Kaplan-Meier Plotter database.Lastly,we examined the infiltration of immune cells in these differentially expressed genes using the TIMER database.Verify the expression level of FAM83A in LUAD cells using real-time fluorescence quantitative PCR(qRT-PCR).Results In GEPIA,the top 10 genes which show differential expression in LUAD are RP11-40C6.2,XAGE1A,XAGE1B,RP5-940J5.9,IGHGP,CRABP2,FAM83A,SPP1,TMPRSS4,and CEACAM5.All these genes have high expression levels in LUAD,except for RP11-40C6.2,XAGE1A,XAGE1B,RP5-940J5.9,CRABP2,SPP1,TMPRSS4,and CEACAM5,which are not significantly correlated with LUAD stage.However,the gene expression levels of IGHGP and FAM83A are correlated with the LUAD stage.In Kaplan-Meier Plotter survival analysis,it was observed that the expression levels of XAGE1A,XAGE1B,CRABP2,FAM83A,SPP1,and TMPRSS4 were negatively correlated with OS.In contrast,XAGE1A,XAGE1B,CRABP2,FAM83A,and SPP1 expression levels were negatively correlated with FP.Additionally,it was found that the expression level of FAM83A was negatively correlated with PPS.In TIMER im-mune cell infiltration analysis,CRABP2 was positively correlated with CD4+T cells,macrophages,neutrophils,and dendritic cells,and negatively correlated with tumor purity.FAM83A was negatively correlated with B cells and dendritic cells,whereas SPP1 was positively correlated with macrophages,neutrophils,and dendritic cells,and negatively correlated with tumor purity.TMPRSS4 was positively cor-related with CD4+T cells and negatively correlated with CD8+T cells,and CEACAM5 was positively correlated with CD4+T cells and negatively correlated with CD8+T cells.The qRT-PCR experiment results showed that FAM83A was highly expressed in LUAD cells.Conclusion Some genes show different levels of expression in LUAD tissue compared to normal tissue.One of these genes is FAM83A,which is highly expressed in lung adenocarcinoma cells and is associated with the stage of the cancer,immune cell infiltra-tion,and poor prognosis in LUAD patients.Therefore,it is possible that FAM83A could be used as a potential biomarker to predict sur-vival prognosis.
基金项目
国家自然科学基金地区基金项目(82360540)
广西医疗卫生适宜技术开发与推广应用重点项目(S2021104)
湖北陈孝平科技发展基金(CXPJJH123003-039)
NETL
NSTL
万方数据