非小细胞肺癌中WASF1基因与免疫细胞浸润的相关性分析

Expression of WASF1 in non-small cell lung cancer and its correlation with tumor-infiltrating immune cells

目的 探讨WASF1在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,及其与NSCLC预后和免疫细胞浸润水平的相关性。方法 分析TCGA数据库中NSCLC和癌旁正常组织中WASF1 mRNA的表达,应用R语言limma分析WASF1 mRNA表达差异。采用TIMER数据库分析NSCLC组织中WASF1 mRNA的表达,采用qRT-PCR和免疫组化法检测NSCLC组织中WASF1 mRNA和蛋白的表达。利用OncoLnc和TIMER数据库分析WASF1对NSCLC患者预后及免疫细胞浸润水平的影响;应用GSEA软件预测WASF1可能参与的信号通路。结果 与癌旁正常组织相比,WASF1在NSCLC组织中高表达,且影响NSCLC患者预后(P<0.05)。肺腺癌中WASF1 mRNA表达与巨噬细胞(r=0.131,P<0.01)、中性粒细胞(r=0.207,P<0.01)和树突状细胞(r=0.093,P<0.05)的免疫浸润水平呈正相关,WASF1的拷贝数变异对B细胞、CD8^(+)T细胞、CD4^(+)T细胞、巨噬细胞、中性粒细胞和树突状细胞的浸润水平均有影响(P<0.05)。B细胞浸润可作为肺腺癌的独立预后因素(HR=0.013,95%CI=0.001~0.187)。肺鳞状细胞癌中WASF1 mRNA表达与细胞纯度呈正相关(r=0.275,P<0.01),与CD8^(+)T细胞(r=-0.138,P<0.01)、CD4^(+)T细胞(r=-0.132,P<0.01)、巨噬细胞(r=-0.149,P<0.01)、中性粒细胞(r=-0.247,P<0.01)和树突状细胞(r=-0.186,P<0.01)的免疫浸润水平呈负相关,WASF1的拷贝数变异对B细胞、CD4^(+)T细胞、中性粒细胞和树突状细胞的浸润水平有影响(P<0.05)。GSEA分析显示WASF1可能通过影响细胞周期、核苷酸切除修复和DNA复制等信号通路,从而导致NSCLC的进展。结论 WASF1在NSCLC中高表达,影响NSCLC预后和肿瘤免疫细胞浸润水平。WASF1可能通过多条通路调控NSCLC的发生、发展。

Purpose To investigate the expression of WASF1 in non-small cell lung cancer(NSCLC) and its correlation with the prognosis and immune cell infiltration level of NSCLC.Methods The mRNA expression levels of WASF1 in NSCLC and normal tissues adjacent to cancer were analyzed in TCGA database,and the mRNA expression differences of WASF1 were analyzed by using R language limma package.The mRNA expression of WASF1 in NSCLC was analyzed by TIMER database.The mRNA and protein expression of WASF1 in NSCLC was detected by qRT-PCR and immunohistochemistry.OncoLnc and TIMER database were used to analyze the effects of WASF1 on prognosis and immune cell infiltration level of NSCLC patients.GSEA software was used to predict the signaling pathways in which WASF1 might be involved.Results WASF1 was highly expressed in NSCLC tissues and affected the prognosis of NSCLC patients(P<0.05).The mRNA expression of WASF1 in LUAD was positively correlated with the level of infiltration of macrophages(r=0.131,P<0.01),neutrophils(r=0.207,P<0.01) and dendritic cells(r=0.093,P<0.05).The infiltration levels of B cells,CD8^(+)T cells,CD4^(+)T cells,macrophages,neutrophils and dendritic cells were affected by the CNV variation of WASF1(P<0.05).B cell infiltration was an independent prognostic factor for lung adenocarcinoma(LUAD)(HR=0.013,95% CI=0.001~0.187).The mRNA expression of WASF1 gene in lung squamous cell carcinoma(LUSC) was positively correlated with cell purity(r=0.275,P<0.01),and negatively correlated with the level of immune infiltration of CD8^(+)T cells(r=-0.138,P<0.01),CD4^(+)T cells(r=-0.132,P<0.01),macrophages(r=-0.149,P<0.01),neutrophils(r=-0.247,P<0.01) and dendritic cells(r=-0.186,P<0.01).The infiltration levels of B cells,CD4^(+)T cells,neutrophils and dendritic cells were affected by the copy number variation of WASF1(P<0.05).GSEA analysis showed that WASF1 may affect the progression of NSCLC by affecting cell cycle,nucleotide excision repair and DNA replication signaling pathways.Conclusion WASF1 is highly expressed in NSCLC and may affect the prognosis of NSCLC patients and the infiltration level of tumor immune infiltrating cells.WASF1 may regulate the occurrence and development of NSCLC through multiple pathways.