肉芽肿性乳腺炎病灶组织差异表达蛋白质筛选及关键分子验证

Identification of differentially expressed proteins of granulomatous mastitis and key cytokine validation

目的分析肉芽肿性小叶性乳腺炎(GLM)病灶组织中的差异表达蛋白质,筛选GLM的生物标志物,再根据结果对关键分子进行验证。方法采用TMT标记定量蛋白质组学方法,对GLM患者(3例)病灶与正常乳腺组织中蛋白质组进行鉴定和定量,筛选出发生显著变化的差异表达蛋白质,导入Proteome Discoverer软件中,搜索Uniprot数据库,完成生物信息学分析。采用细胞因子抗体芯片技术对筛选出的关键分子进行定量和验证。结果GLM病灶组织与正常乳腺组织筛选出差异表达蛋白质1405个,其中1106个表达上调,299个表达下调。GO富集分析显示,这些差异表达蛋白质主要参与细胞过程、代谢过程、蛋白质代谢、生物学调节等。KEGG通路分析发现,差异表达蛋白质参与补体活化、免疫调节、炎症反应等。细胞因子定量分析验证了GLM病灶组织中IL-1β、MCP2、IL-6表达升高。结论GLM病灶和正常乳腺组织中存在差异表达的蛋白质参与炎症、免疫等多种信号通路,可能是GLM发生发展的关键分子。特定蛋白质是值得进一步研究的生物标志物。

Objective To identify the differentially expressed proteins in patients of granulomatous lobular mastitis(GLM)and screen the biomarkers that related to GLM,then to verify some key molecule according to the results.Methods The lesion tissues and adjacent normal tissues following surgical removal of affected tissues were collected from 3 GLM patients.Then the quantitative proteomics TMT technology was used to detect and quantitatively analyze the differentially expressed proteins,and to conduct functional analysis.Uniprot database and proteome discoverer software were applied to analyze the bioinformatic.The concentrations of key cytokine were analyzed by human antibody array.Results In this study,1405 differentially expressed proteins were identified including 1106 upregulated proteins and 299 downregulated proteins in GLM.Gene ontology(GO)annotation and functional cluster analyses showed that these differentially expressed proteins mainly involved in cellular processes,metabolic processes,protein metabolism,biological regulation.KEGG pathway analysis showed these differentially expressed proteins were mainly enriched in complement activation,immune system,inflammatory response.Cytokine quantitative analysis confirmed the increased expression of IL-1β,MCP2 and IL-6 in GLM tissures.Conclusion Differentially expressed proteins in granulomatous mastitis are involved in related signaling pathways and may be key molecules in the development of granulomatous mastitis.Specific proteins are the potential biomarkers that deserve further exploration.