Identification of key immune cell-related genes involved in tumorigenesis and prognosis of cervical squamous cell carcinoma

The infiltration of immune cells can significantly affect the prognosis and immune therapy of patients with cervical squamous cell carcinoma (CSCC). This study aimed to explore key immune cell-related genes in the tumorigenesis and prognosis of CSCC. The module significantly related to immunity was screened by weighted gene co-expression network analysis (WGCNA) and ESTIMATE analysis, followed by correlation analysis with clinical traits. Key candidate genes were intersected with the protein–protein interaction (PPI) network genes for immune-related genes. The relationship between immune cell infiltration and key genes was analyzed. Tumor immune dysfunction and exclusion (TIDE) and immunophenoscore (IPS) predicted the response to immunotherapy in CSCC patients. Clinically, quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry were manipulated for analyzing the changes in mRNA and protein expression of key genes in cancer. Western blot was conducted to assess the correlation between key genes and immune infiltration. The brown module was notably associated with the immune microenvironment of CSCC, from which three immune-related key genes (<i>TYROBP</i>, <i>CCL5</i>, and <i>HLA-DRA</i>) were obtained. High expression of these genes was significantly positively associated with the infiltration abundance of T cells, B cells, and other immune cells. High expression levels of three key genes were confirmed in para-cancer tissue and correlated with the abundance of immune cells. The high-expression group of key genes was more sensitive to immunotherapy. We provide a theoretical basis for searching for potential targets for effective treatment and diagnosis of CSCC and provide new ideas for developing novel immunotherapy strategies.

免疫细胞浸润可显著影响宫颈鳞状细胞癌（CSCC）患者的预后与免疫治疗效果。本研究旨在探索宫颈鳞状细胞癌发生发展及预后过程中的关键免疫细胞相关基因。通过加权基因共表达网络分析（WGCNA）与ESTIMATE分析筛选出与免疫显著相关的模块，随后结合临床特征开展关联分析。将关键候选基因与免疫相关基因的蛋白质相互作用（PPI）网络基因取交集，分析免疫细胞浸润与关键基因之间的关联。采用肿瘤免疫功能异常与排斥（TIDE）评分与免疫表型评分（IPS）预测宫颈鳞状细胞癌患者的免疫治疗响应情况。临床层面，通过实时定量聚合酶链反应（qRT-PCR）与免疫组化实验分析关键基因在癌组织中的mRNA与蛋白表达变化；采用蛋白质印迹法（Western blot）评估关键基因与免疫浸润的相关性。研究发现，棕色模块与宫颈鳞状细胞癌的免疫微环境显著相关，从中筛选得到3个免疫相关关键基因：<i>TYROBP</i>、<i>CCL5</i>及<i>HLA-DRA</i>。上述基因的高表达与T细胞、B细胞及其他免疫细胞的浸润丰度呈显著正相关。本研究在癌旁组织中验证了这3个关键基因的高表达水平，且其表达水平与免疫细胞丰度相关。关键基因高表达组患者对免疫治疗更为敏感。本研究可为宫颈鳞状细胞癌的有效治疗与诊断潜在靶点的筛选提供理论依据，同时为新型免疫治疗策略的开发提供新思路。