鉴定代谢蛋白ATP5MF为三阴性乳腺癌(TNBC)潜在的治疗靶点

三阴性乳腺癌(TNBC)作为乳腺癌的一个特殊亚型，其特征为高侵袭性、高转移潜能、易复发和预后不良。针对非BRCA1/2突变TNBC的有效治疗方案仍然缺乏，因此迫切需要开发新的治疗策略。本研究中，单细胞RNA测序数据来自GEO数据库，转录组数据则来自TCGA和METABRIC数据库。对单细胞测序数据进行了质量控制，随后采用注释和Copycat算法进行分析。利用高维加权基因共表达网络分析（hdWGCNA）方法，分析非BRCA1/2突变TNBC的肿瘤上皮细胞，鉴定功能模块基因。通过蛋白质相互作用（PPI）分析和生存分析，进一步筛选出关键基因。将siRNA-NC和siRNA-ATP5MF转染入两株MDA-MB-231和BT-549TNBC细胞系，利用CCK8法、克隆形成和迁移实验检测细胞生长。电子显微镜观察细胞线粒体结构，JC-1染色测定线粒体膜电位。在裸鼠中建立肿瘤异种移植模型，评估ATP5MF沉默后体内肿瘤反应。通过hdWGCNA，我们在模块3中鉴定出136个基因，经PPI和生存分析筛选出ATP5MF作为潜在治疗基因。ATP5MF高表达与非BRCA1/2突变TNBC患者预后不良相关。在TCGA数据库及临床TNBC标本免疫组化染色中检测ATP5MF在TNBC组织中的高表达。沉默ATP5MF显著抑制体外TNBC细胞的生长和克隆形成，阻碍TNBC异种移植物的生长，并损害TNBC细胞的线粒体功能。综上所述，代谢蛋白ATP5MF在非BRCA1/2突变TNBC细胞中发挥关键作用，具有潜在的诊断和治疗新靶点价值。

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.