卵巢癌关键基因CLDN4的风险模型构建与免疫细胞浸润分析

Risk model construction and immune cell infiltration analysis of the CLDN4 gene in ovarian cancer cells

目的筛选卵巢癌(OV)发生过程中的关键基因,并进行免疫细胞浸润分析和风险模型构建,为OV患者的早期诊治和预后评估提供依据。方法首先,通过数据分析工具(GEO2R)对GSE18520和GSE6008数据进行差异表达基因(DEG)筛选并绘制韦恩图;其次,对DEG进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)、蛋白互作(PPI)网络、突变与表达和预后分析,从而确定关键基因;再次,对关键基因进行风险模型构建与免疫细胞浸润分析;最后,临床收集OV组织作为实验组,收集癌旁正常组织作为对照组,通过实时定量PCR和Western blot法检测两组中紧密连接蛋白4(CLDN4)mRNA和蛋白的表达情况,并对两组结果进行对比分析。结果CLDN4是OV发生过程中的关键基因,且随着其表达的增高,OV患者的预后风险增加,不利于患者的总体生存(OS)。CLDN4与OV微环境中的树突状细胞(DC)显著正相关,且高表达的DC对OV患者OS显著有利。CLDN4的mRNA在OV组织中表达量显著升高,CLDN4的蛋白表达水平在OV组织中显著增高,差异均具有统计学意义。结论CLDN4是OV发生过程中的关键基因,可作为OV的潜在生物标志物和免疫治疗靶点。

Objective To screen for the key genes involved in the development of ovarian cancer(OV),analyze the immune cell infiltration and construct a risk model,so as to provide evidence for the early diagnosis,treatment and prognosis evaluation of OV patients.Methods The GSE18520 and GSE6008 datasets were analyzed for differentially expressed genes(DEGs)using the GEO2R data analysis tool,and a Venn diagram was generated.Then,DEGs were subjected to Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,and protein-protein interaction(PPI)networks,as well as mutations,expression and prognosis analysis to identify key genes.Next,a risk model was constructed and immune cell infiltration analysis of key genes was performed.Finally,ovarian cancer tissues were collected as the experimental group,and adjacent normal tissues were collected as the control group.The expression of claudin 4(CLDN4)mRNA and protein levels were detected using real-time quantitative PCR(RT-qPCR)and Western-blot,and the results were compared between the two groups.Results CLDN4 was identified as a key gene in the development of OV.As its expression increased,the prognosis risk of OV patients worsened,which unfavorably impacted their overall survival(OS).A significant positive correlation was found between CLDN4 and dendritic cell(DC)in the OV microenvironment,and high expression of DCs was significantly associated with better OS in OV patients.The mRNA and protein expression levels of CLDN4 were significantly increased in OV tissues,with statistically significant differences.Conclusion CLDN4 is a key gene in the development of OV,and may serve as a potential biomarker and immunotherapy target for OV.