三阴性乳腺癌（TNBC）是乳腺癌的一种独特亚型，具有高侵袭性、高转移性、易复发、预后差等特点。非BRCA1/2突变TNBC仍缺乏有效的治疗方案。因此，临床迫切需要针对非BRCA1/2突变的TNBC开发新的治疗方法。本研究的scRNA数据来自GEO数据库，转录组数据来自TCGA和METABRIC数据库。对单细胞测序数据进行质量控制程序。然后应用注释和Copycat算法进行分析。采用高维加权基因共表达网络分析（hdWGCNA）方法，我们分析了非 BRCA1/2 突变 TNBC 的肿瘤上皮细胞，以确定功能模块基因。进一步利用PPI分析和生存分析来鉴定关键基因。将 siRNA-NC 和 siRNA-ATP5MF 转染至两种 MDA-MB-231 和 BT-549 TNBC 细胞系中。通过CCK8测定、集落形成和迁移测定来测定细胞生长。电子显微镜用于检查细胞中线粒体的结构。 JC-1染色用于测量线粒体膜的电位。通过将TNBC细胞注射到裸鼠体内建立肿瘤异种移植动物模型。该动物模型用于评估ATP5MF沉默后的体内肿瘤反应。使用hdWGCNA，我们在模块3中鉴定了136个基因。经过PPI和生存分析，我们鉴定了ATP5MF作为潜在的治疗基因。 ATP5MF 高表达与非 BRCA1/2 突变 TNBC 的不良预后相关。使用TCGA数据库和临床TNBC标本的IHC染色评估TNBC组织中ATP5MF的高表达。在两种TNBC细胞系中沉默ATP5MF可减少体外TNBC细胞的生长和集落形成，并阻碍体内TNBC异种移植物的生长。此外，ATP5MF敲低会损害TNBC细胞中的线粒体功能。 总之，代谢蛋白ATP5MF在非BRCA1/2突变TNBC细胞中发挥着至关重要的作用，使其成为非BRCA1/2突变TNBC潜在的新型诊断和治疗肿瘤靶点.© 2024。作者。

Triple-negative breast cancer (TNBC), a distinct subtype of breast cancer, is characterized by its high invasiveness, high metastatic potential, proneness to relapse, and poor prognosis. Effective treatment regimens for non-BRCA1/2 mutation TNBC are still lacking. As a result, there is a pressing clinical necessity to develop novel treatment approaches for non-BRCA1/2 mutation TNBC.For this research, the scRNA data was obtained from the GEO database, while the transcriptome data was obtained from the TCGA and METABRIC databases. Quality control procedures were conducted on single-cell sequencing data. and then annotation and the Copycat algorithm were applied for anlysis. Employing the high dimensional weighted gene coexpression network analysis (hdWGCNA) method, we analyzed the tumor epithelial cells from non-BRCA1/2 mutation TNBC to identify the functional module genes. PPI analysis and survival analysis were further emplyed to identify the key gene. siRNA-NC and siRNA-ATP5MF were transfected into two MDA-MB-231 and BT-549 TNBC cell lines. Cell growth was determined by CCK8 assay, colony formation and migration assay. Electron microscopy was used to examine the structure of mitochondria in cells. JC-1 staining was used to measure the potential of the mitochondrial membrane. A tumor xenograft animal model was established by injecting TNBC cells into nude mice. The animal model was usded to evaluated in vivo tumor response aftering ATP5MF silencing.Using hdWGCNA, we have identified 136 genes in module 3. After PPI and survival analysis, we have identified ATP5MF as a potential therapeutic gene. High ATP5MF expression was associated with poor prognosis of non-BRCA1/2 mutation TNBC. The high expression of ATP5MF in TNBC tissues was evaluated using the TCGA database and IHC staining of clinical TNBC specimens. Silencing ATP5MF in two TNBC cell lines reduced the growth and colony formation of TNBC cells in vitro, and hindered the growth of TNBC xenografts in vivo. Additionally, ATP5MF knockdown impaired mitochondrial functions in TNBC cells.In summary, the metabolic protein ATP5MF plays a crucial role in the non-BRCA1/2 mutation TNBC cells, making it a potential novel diagnostic and therapeutic oncotarget for non-BRCA1/2 mutation TNBC.© 2024. The Author(s).