铁死亡是一种铁依赖形式的程序性细胞死亡，对癌症有显着影响，但其与前列腺癌 (PCa) 预后的联系仍未得到充分研究。本研究旨在开发和验证铁死亡相关基因特征，以预测 PCa 预后和免疫微环境差异，从而潜在地确定治疗靶点。478 名 PCa 患者的 RNA 测序数据和相应的临床数据从癌症基因组图谱 (TCGA) 数据库下载。我们使用 Kaplan-Meier 方法调查了高风险组和低风险组的无病生存率 (DFS)。通过基因集富集分析（GSEA）、基因本体论（GO）和京都基因和基因组百科全书（KEGG）分析来调查高风险组和低风险组之间的功能差异。使用 CIBERSORT 检查铁死亡风险评分和免疫状态之间的联系。使用定量实时聚合酶链反应 (qRT-PCR)、蛋白质印迹和免疫组织化学 (IHC) 验证良性前列腺增生 (BPH) 和 PCa 中核心预后基因的表达水平。一种新的铁死亡相关预后基因特征是基于单变量和多变量 Cox 回归分析在基因表达综合 (GEO) 数据库中建立和测试。根据这种铁死亡特征，PCa 患者被分为高风险组和低风险组。高风险组患者的预后比低风险组更差。该模型的预测准确性通过受试者工作特征 (ROC) 分析得到证明。 TCGA 队列的另一项富集分析显示，高危组中免疫相关通路显着上调，1 年时曲线下面积 (AUC) 为 0.85，3 年时为 0.82，5 年时为 0.76。在 GEO 队列中，AUC 在 1 年时达到 0.69，在 3 年时达到 0.74，在 5 年时达到 0.75。另一项富集分析表明，高危人群中细胞因子相关途径、免疫受体活性和其他免疫相关途径显着上调。此外，分析显示，与低危组PCa患者相比，高危组肥大细胞和浆细胞的比例显着较低。相反，高风险组中调节性T细胞（Treg）的比例显着高于低风险组。根据 qRT-PCR、Western blot 和 IHC 结果，DRD4、SRC、AKR1C2 和 AIFM2 在 PCa 中的表达显着高于 BPH。我们还表明，铁他汀 1 处理的 LNCaP 细胞具有较高的 DRD4、SRC 和 AKR1C2 表达水平。构建并验证了八个铁死亡相关基因 (FRG) 的预后特征，可以准确预测 PCa 患者的结果。 FRGs 可能有助于抗肿瘤免疫并作为 PCa.2024 AME Publishing Company 的治疗靶点。版权所有。

Ferroptosis, an iron-dependent form of programmed cell death, significantly impacts cancer, yet its link to prostate cancer (PCa) prognosis remains underexplored. This study aims to develop and validate a ferroptosis-related gene signature to predict PCa prognosis and immune microenvironment differences, potentially identifying therapeutic targets.RNA-sequencing data of 478 PCa patients and corresponding clinical data were downloaded from The Cancer Genome Atlas (TCGA) database. We investigated the disease-free survival (DFS) rates of the high- and low-risk groups using the Kaplan-Meier method. Functional differences between the high- and low-risk groups were investigated by a gene set enrichment analysis (GSEA), and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The link between ferroptosis risk score and immune status was examined using CIBERSORT. The expression levels of core prognostic genes in benign prostatic hyperplasia (BPH) and PCa were verified using quantitative real-time polymerase chain reaction (qRT-PCR), Western blot, and immunohistochemistry (IHC).A novel ferroptosis-related prognostic gene signature was established and tested in the Gene Expression Omnibus (GEO) database based on univariate and multivariate Cox regression analyses. Patients with PCa were classified into high- and low-risk groups based on this ferroptosis signature. Patients in the high-risk group had worse outcomes than those in the low-risk group. The predictive accuracy of the model was demonstrated by a receiver operating characteristic (ROC) analysis. An additional enrichment analysis of TCGA cohort revealed the immune-related pathways were significantly upregulated in the high-risk group, with areas under the curve (AUCs) of 0.85 at 1 year, 0.82 at 3 years, and 0.76 at 5 years. In the GEO cohort, the AUCs reached 0.69 at 1 year, 0.74 at 3 years, and 0.75 at 5 years. An additional enrichment analysis indicated a significant upregulation of cytokine-related pathways, immune receptor activity, and other immune-related pathways in the high-risk group. Furthermore, the analysis revealed that the proportions of mast cells and plasma cells were significantly lower in the high-risk group compared to the low-risk group of PCa patients. Conversely, the proportion of regulatory T cells (Tregs) was significantly higher in the high-risk group than in the low-risk group. According to the qRT-PCR, Western blot, and IHC results, DRD4, SRC, AKR1C2, and AIFM2 expression was significantly higher in PCa than BPH. We also showed that the ferrostatin 1-treated LNCaP cells had higher expression levels of DRD4, SRC, and AKR1C2.A prognostic signature of eight ferroptosis-related genes (FRGs) that may accurately predict PCa patient outcomes was constructed and validated. FRGs may contribute to anti-tumor immunity and serve as therapeutic targets in PCa.2024 AME Publishing Company. All rights reserved.