NUSAP1在食管癌中与肿瘤细胞增殖和免疫细胞浸润相关性——基于数据库的研究

Association of NUSAP1 with tumor cell proliferation and immune cell infiltration in esophageal carcinoma: database based research

摘要:

目的 探讨核仁纺锤体相关蛋白1（nucleolar and spindle-associated protein 1，NUSAP1）在食管癌（esophageal carcinoma，ESCA）中的表达情况、临床意义及功能机制，为ESCA的早期诊断和治疗提供新思路。方法 从GEPIA2和TIMER数据库下载ESCA样本与癌旁样本，进行NUSAP1的表达分析，STRING和Cytoscape软件进行NUSAP1相关基因的PPI网络构建，并进行GO、KEGG及GSEA富集分析。TIMER数据库被用来分析NUSAP1与免疫细胞的相关性。最后通过RT-PCR、Western blot和免疫组化验证NUSAP1在ESCA组织和细胞中的表达，使用慢病毒包被的小干扰RNA转染ESCA细胞，进行NUSAP1的表达敲减。通过CKK-8和Ki67免疫荧光评估NUSAP1对ESCA细胞增殖的影响。结果 NUSAP1在ESCA组织和细胞中高表达。NUSAP1的表达与CDK1、B细胞和巨噬细胞浸润正相关，而与CD4⁺ T细胞、CD8⁺ T细胞、中性粒细胞和树突状细胞浸润负相关。GSEA分析发现NUSAP1的高表达组主要富集于Wnt/β-catenin 信号通路、过氧化物酶通路、PI3K/Akt/mTOR信号通路。在体外，敲减NUSAP1后抑制了ESCA细胞增殖。结论 NUSAP1在ESCA中高表达，并与ESCA细胞增殖和免疫细胞浸润密切相关。因此，NUSAP1可能有助于ESCA的诊断和治疗。

Abstract:

Objective To investigate the expression，clinical significance and functional mechanism of nucleolar and spindle-associated protein 1（NUSAP1）in esophageal carcinoma（ESCA），and to provide new ideas for the early diagnosis and treatment of ESCA.Methods ESCA samples and paracancerous samples were downloaded from GEPIA2 and TIMER databases to analyze the expression of NUSAP1，STRING and Cytoscape software were used to construct the PPI network of NUSAP1 related genes，and GO，KEGG and GSEA enrichment analysis were performed. The TIMER database was used to analyze the correlation between NUSAP1 and immune cells. Finally，the expression of NUSAP1 in ESCA tissues and cells was verified by RT-PCR，Western blot and immunohistochemistry，and ESCA cells were transfected with lentivirus-coated small interfering RNA to knock down the expression of NUSAP1. The effect of NUSAP1 on ESCA cell proliferation was evaluated by CKK-8 and Ki67 immunofluorescence.Results NUSAP1 was highly expressed in ESCA tissues and cells. The expression of NUSAP1 was positively correlated with the infiltration of CDK1，B cells and macrophages，but negatively correlated with the infiltration of CD4⁺ T cells，CD8⁺ T cells，neutrophils and dendritic cells. GSEA analysis found that the high expression group of NUSAP1 was mainly enriched in Wnt/β-catenin signaling pathway，peroxidase pathway，PI3K/Akt/mTOR signaling pathway. In vitro，knockdown of NUSAP1 inhibited the proliferation of ESCA cells.Conclusion NUSAP1 is highly expressed in ESCA and closely related to ESCA cell proliferation and immune cell infiltration. Therefore，NUSAP1 may help the diagnosis and treatment of ESCA.