【作者】 周湘；

【导师】 刘红光；

【作者基本信息】 南华大学 ， 临床医学（外科学）（专业学位）， 2024， 硕士

【摘要】 目的:探究SLC40A1在乳腺癌组织的表达水平及临床预后的相关性,同时分析该基因在肿瘤微环境中的作用和肿瘤细胞增殖的相关性。方法:1.从TCGA数据库中下载乳腺癌患者的RNA-seq表达谱和相应的临床数据,并对其进行免疫细胞相关性分析。2.提取免疫及代谢相关的基因表达数据集,并联合上述乳腺癌mRNA三者取交集建立与免疫及代谢相关的乳腺癌mRNA基因表达数据集,并进行表达差异分析获取差异基因和KEGG与GO富集分析。3.通过单因素COX回归分析及结合文献筛选出SLC40A1基因作为目标基因。4.利用R语言对目标基因进行表达差异、生存、与临床特征的关系、单基因GSEA富集分析和免疫浸润分析。5.通过在线网络数据库进行验证、补充分析并分析其与肿瘤微环境细胞成分进行相关性分析等。6.通过免疫组化染色法检测乳腺癌和癌旁正常组织中SLC40A1基因的表达差异,不同表达水平下与临床特征的关系以及SLC40A1基因表达水平与增殖指标Ki67的关系。结果:1)SLC40A1在乳腺癌及癌旁表达水平有差异,乳腺癌表达水平低于癌旁组织;2)SLC40A1低水平乳腺癌患者预后差,SLC40A1低水平与CD8+T Cell浸润相关,提示预后差;3)SLC40A1高表达乳腺癌样本往往伴随着较高的StromalScore和ESTIMATEScore,而SLC40A1低表达的乳腺癌样本则具有较低的StromalScore和ESTIMATEScore;4)SLC40A1表达与乳腺癌肿瘤微环境中CD8+T Cell、巨噬细胞和肿瘤相关纤维细胞等成正相关,与乳腺癌增殖相关基因成负相关;5)SLC40A1在不同临床特征(T、N、M分期和分子受体表达情况)的乳腺癌中具有表达水平差异,临床分期越晚,SLC40A1表达水平越低,激素受体阳性患者该基因表达水平高于激素受体阴性;6)SLC40A1高表达与乳腺癌患者的治疗反应较差相关。7)SLC40A1表达水平在乳腺癌不同病理分级、淋巴结情况、ER表达状态和Ki67百分比有差异;8)SLC40A1表达水平与乳腺癌Ki67呈负相关。结论:1.在乳腺癌中,SLC40A1基因表达普遍较低,可能与预后不良有关;2.SLC40A1表达可能与Ki67呈负相关,揭示其可能与细胞增殖相关。更多还原

【Abstract】 Purpose: This study aims to investigate the expression levels of SLC40A1 in breast cancer tissue and its correlation with clinical prognosis.Additionally,we analyze the role of this gene in the tumor microenvironment and its association with tumor cell proliferation.Methods:1.RNA-seq expression profiles and corresponding clinical data of breast cancer patients were downloaded from the TCGA database,and immune cell-related correlation analysis was conducted.2.Immune and metabolism-related gene expression datasets were extracted,and the intersection of these datasets with breast cancer mRNA was used to establish an immune and metabolism-related gene expression dataset.Differential expression analysis was performed to obtain differentially expressed genes,followed by KEGG and GO enrichment analysis.3.SLC40A1 gene was selected as the target gene through univariate COX regression analysis and literature screening.4.R language was used for differential expression analysis,survival analysis,analysis of the relationship between the target gene and clinical features,single-gene GSEA enrichment analysis,and immune infiltration analysis.5.Online databases were utilized for validation,supplementary analysis,and correlation analysis with tumor microenvironment cell components.6.Immunohistochemistry staining was performed to detect the expression difference of SLC40A1 gene in breast cancer and adjacent normal tissues,as well as its relationship with clinical features and Ki67,a proliferation marker.Results:1)SLC40A1 showed differential expression levels between breast cancer and adjacent tissues,with lower expression in breast cancer tissues.2)Breast cancer patients with low levels of SLC40A1 had worse prognosis,and low SLC40A1 expression was associated with CD8+ T cell infiltration,indicating poor prognosis.3)Breast cancer samples with high SLC40A1 expression were often accompanied by higher StromalScore and ESTIMATEScore,while breast cancer samples with low SLC40A1 expression had lower StromalScore and ESTIMATEScore.4)SLC40A1 expression was positively correlated with CD8+ T cells,macrophages,and tumor-associated fibroblasts in the breast cancer tumor microenvironment,and negatively correlated with genes related to tumor proliferation.5)SLC40A1 showed differential expression levels in breast cancer with different clinical features(T,N,M staging,and molecular receptor expression).The expression level of SLC40A1 decreased as the clinical stage advanced,and hormone receptor-positive patients had higher expression levels compared to hormone receptor-negative patients.6)High expression of SLC40A1 was associated with poorer treatment response in breast cancer patients.7)SLC40A1 expression levels varied in breast cancer based on different pathological grades,lymph node status,ER expression status,and Ki67 percentage.8)SLC40A1 expression levels were negatively correlated with Ki67 in breast cancer.Conclusion:1.In breast cancer,SLC40A1 gene expression is generally low,suggesting a potential association with poor prognosis.2.SLC40A1 expression may be negatively correlated with Ki67,indicating its potential involvement in cell proliferation.更多还原