Department of General Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, Jiangsu, China.
The Graduate School of Nanjing Medical University, Nanjing, Jiangsu, China.
J Transl Med. 2024 Oct 14;22(1):932. doi: 10.1186/s12967-024-05692-9.
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.
三阴性乳腺癌（TNBC）是一种独特的乳腺癌亚型，具有侵袭性高、转移潜能高、易复发和预后差等特点。对于非 BRCA1/2 突变的 TNBC，目前仍缺乏有效的治疗方案。因此，迫切需要为非 BRCA1/2 突变的 TNBC 开发新的治疗方法。
本研究从 GEO 数据库中获取 scRNA 数据，从 TCGA 和 METABRIC 数据库中获取转录组数据。对单细胞测序数据进行质量控制处理，然后进行注释和 Copycat 算法分析。采用高维加权基因共表达网络分析（hdWGCNA）方法，分析非 BRCA1/2 突变的 TNBC 肿瘤上皮细胞，鉴定功能模块基因。进一步进行 PPI 分析和生存分析，鉴定关键基因。将 siRNA-NC 和 siRNA-ATP5MF 转染到 MDA-MB-231 和 BT-549 两种 TNBC 细胞系中。通过 CCK8 测定、集落形成和迁移测定来确定细胞的生长情况。使用电子显微镜观察细胞中线粒体的结构。使用 JC-1 染色来测量线粒体膜的电位。通过将 TNBC 细胞注入裸鼠建立肿瘤异种移植动物模型。使用该动物模型评估沉默 ATP5MF 后体内肿瘤的反应。
通过 hdWGCNA，我们在模块 3 中鉴定了 136 个基因。经过 PPI 和生存分析，我们发现 ATP5MF 是一个潜在的治疗靶点。高 ATP5MF 表达与非 BRCA1/2 突变的 TNBC 患者的不良预后相关。通过 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 可降低 TNBC 细胞的体外生长和集落形成，并抑制 TNBC 异种移植瘤在体内的生长。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 突变的 TNBC 细胞中发挥着关键作用，使其成为非 BRCA1/2 突变的 TNBC 的一种潜在的新型诊断和治疗靶点。
