Zhang Kun, Xie Huyang, Zhao Fan, Huang Yeqing
Department of Urology, Affiliated Hospital of Nantong University, Nantong, China.
Front Immunol. 2025 Apr 16;16:1564784. doi: 10.3389/fimmu.2025.1564784. eCollection 2025.
Prostate adenocarcinoma (PRAD) is a leading cause of male mortality, with NK/T cell communication being key areas of the research.
The Seurat package was utilized to normalize and reduce the dimensionality of the single-cell data, and CellMarker 2.0 was employed for cell annotation. CellChat was utilized to construct the ligand-receptor interaction network of cell subsets. Differentially expressed genes (DEGs) were filtered by the limma package. Univariate Cox and the LASSO regression in the glmnet package were used to obtain biomarkers and develop a risk model. The survminer package was used to calculate the optimal threshold for dividing patients into high-risk and low-risk groups, and then Kaplan-Meier (KM) survival analysis was performed. Single-sample GSEA (ssGSEA), TIMER, and ESTIMATE packages were employed for immune infiltration analysis. Pathway analysis was conducted for the low- and high-risk groups using GSEA. Immunotherapy responses were evaluated by adopting TIDE method. Additional cellular validation (quantitative real-time PCR, CCK-8, Transwell, and scratch assay) was implemented to confirm the effects of feature genes on PRAD.
Compared with the benign group, NK/T cells were the cell type with the greatest changes in the tumor group, and their communication intensity was relatively high among all cell types. A RiskScore model was constructed as follows: . Analysis of the differences between the two risk groups showed that the level of immune infiltration was higher in the high-risk group, and it was significantly enriched in immune-correlated pathways, while the low-risk group was mainly enriched in metabolism-related pathways. TIDE analysis indicated that the high-risk group had higher immune escape potential. The cellular validation assays have revealed the higher expression of seven biomarkers in PRAD groups. Further, knockdown inhibited the proliferation, migration, and invasion ability of PRAD cells.
The current research reveals key communication genes in PRAD, offering new possibilities for the exploration of new therapeutic targets.
前列腺腺癌（PRAD）是男性死亡的主要原因，自然杀伤细胞（NK）/T细胞通讯是研究的关键领域。
利用Seurat软件包对单细胞数据进行标准化和降维处理，并使用CellMarker 2.0进行细胞注释。利用CellChat构建细胞亚群的配体-受体相互作用网络。通过limma软件包筛选差异表达基因（DEG）。使用单变量Cox回归和glmnet软件包中的LASSO回归来获得生物标志物并建立风险模型。使用survminer软件包计算将患者分为高危和低危组的最佳阈值，然后进行Kaplan-Meier（KM）生存分析。采用单样本基因集富集分析（ssGSEA）、TIMER和ESTIMATE软件包进行免疫浸润分析。使用基因集富集分析（GSEA）对低危和高危组进行通路分析。采用肿瘤免疫逃逸分析（TIDE）方法评估免疫治疗反应。进行了额外的细胞验证（定量实时PCR、CCK-8、Transwell和划痕试验）以确认特征基因对PRAD的影响。
与良性组相比，NK/T细胞是肿瘤组中变化最大的细胞类型，并且在所有细胞类型中它们的通讯强度相对较高。构建了如下风险评分模型： 。对两个风险组之间差异的分析表明，高危组的免疫浸润水平较高，并且在免疫相关通路中显著富集，而低危组主要富集在代谢相关通路中。TIDE分析表明高危组具有更高的免疫逃逸潜力。细胞验证试验显示PRAD组中有七种生物标志物的表达较高。此外， 基因敲低抑制了PRAD细胞的增殖、迁移和侵袭能力。
当前研究揭示了PRAD中的关键通讯基因，为探索新的治疗靶点提供了新的可能性。
