■前列腺癌(PCa)已成为世界上男性发病率最高的恶性肿瘤。肿瘤微环境(TME)是肿瘤生长所必需的。M2巨噬细胞在许多实体瘤中发挥重要作用。本研究旨在探讨M2巨噬细胞在PCa预后中的作用。
■从基因表达综合数据库(GEO)和癌症基因组图谱(TCGA)获得单细胞RNA-seq(scRNA-seq)数据和mRNA表达数据。质量控制,归一化,reduction,聚类,和scRNA-seq数据的细胞注释使用Seruat包进行。分析肿瘤相关巨噬细胞(TAM)的亚群,并选择M2巨噬细胞的标记基因。使用limma鉴定PCa中的差异表达基因(DEGs),并使用CIBERSORTx检测免疫浸润。然后,构建了加权相关网络分析(WGCNA)来鉴定M2巨噬细胞相关模块和基因.整合来自scRNA-seq数据分析的M2巨噬细胞的标记基因和来自WGCNA的hub基因以基于单变量和LASSO回归分析选择预后基因签名。计算了风险评分,还有DEG,生物学功能,探讨了与风险评分相关的免疫特征。并构建了预测列线图。CCK8,Transwell,和伤口愈合用于验证共培养后细胞表型的变化。
■基于scRNA-seq数据和WGCNA总共选择了2431个M2巨噬细胞标记基因和650个hubM2巨噬细胞相关基因。然后,通过重叠scRNA-seq数据和WGCNA结果获得113个M2巨噬细胞相关基因。9个M2巨噬细胞相关基因(SMOC2,PLPP1,HES1,STMN1,GPR160,ABCG1,MAZ,MYC,和EPCAM)被筛选为预后基因特征。计算M2风险评分,DEGs,免疫评分,基质评分,估计得分,肿瘤纯度,免疫细胞浸润,免疫检查点表达,并确定了免疫疗法和化疗的反应。并构建了预测列线图。CCK8Transwell入侵,和伤口愈合进一步验证了M2巨噬细胞促进增殖,入侵,和PCa的迁移(p<0.05)。
■我们发现M2巨噬细胞和相关基因在促进其发生中起关键作用,发展,和PCa的转移,并作为PCa的令人信服的预测因子。
UNASSIGNED: Prostate cancer (PCa) has become the highest incidence of malignant tumor among men in the world. Tumor microenvironment (TME) is necessary for tumor growth. M2 macrophages play an important role in many solid tumors. This research aimed at the role of M2 macrophages\' prognosis value in PCa.
UNASSIGNED: Single-cell RNA-seq (scRNA-seq) data and mRNA expression data were obtained from the Gene Expression Omnibus database (GEO) and The Cancer Genome Atlas (TCGA). Quality control, normalization, reduction, clustering, and cell annotation of scRNA-seq data were preformed using the Seruat package. The sub-populations of the tumor-associated macrophages (TAMs) were analysis and the marker genes of M2 macrophage were selected. Differentially expressed genes (DEGs) in PCa were identified using limma and the immune infiltration was detected using CIBERSORTx. Then, a weighted correlation network analysis (WGCNA) was constructed to identify the M2 macrophage-related modules and genes. Integration of the marker genes of M2 macrophage from scRNA-seq data analysis and hub genes from WGCNA to select the prognostic gene signature based on Univariate and LASSO regression analysis. The risk score was calculated, and the DEGs, biological function, immune characteristics related to risk score were explored. And a predictive nomogram was constructed. CCK8, Transwell, and wound healing were used to verify cell phenotype changes after co-cultured.
UNASSIGNED: A total of 2431 marker genes of M2 macrophage and 650 hub M2 macrophage-related genes were selected based on scRNA-seq data and WGCNA. Then, 113 M2 macrophage-related genes were obtained by overlapping the scRNA-seq data and WGCNA results. Nine M2 macrophage-related genes (SMOC2, PLPP1, HES1, STMN1, GPR160, ABCG1, MAZ, MYC, and EPCAM) were screened as prognostic gene signatures. M2 risk score was calculated, the DEGs, Immune score, stromal score, ESTIMATE score, tumor purity, and immune cell infiltration, immune checkpoint expression, and responses of immunotherapy and chemotherapy were identified. And a predictive nomogram was constructed. CCK8, Transwell invasion, and wound healing further verified that M2 macrophages promoted the proliferation, invasion, and migration of PCa (p < 0.05).
UNASSIGNED: We uncovered that M2 macrophages and relevant genes played key roles in promoting the occurrence, development, and metastases of PCa and played as convincing predictors in PCa.