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目的:分析有腋窝淋巴结转移和无腋窝淋巴结转移的乳腺浸润性导管癌患者血清中的差异蛋白,筛选与乳腺癌转移相关的生物标志物和治疗靶点。方法:本研究采用定量蛋白质组学串联质谱标签(tandem mass tag,TMT)标记技术分别对14例有腋窝淋巴结转移及14例无腋窝淋巴结转移的乳腺癌患者血清蛋白进行检测,并对蛋白质进行定量分析,筛选出发生显著变化的差异蛋白,再搜索Uniprot数据库和用Proteome Discoverer软件分析,进一步进行生物信息学分析。结果:有腋窝淋巴结转移组中共筛选出差异蛋白119个,其中6个表达上调,113个表达下调。基因本体(gene ontology,GO)注释分析和功能聚类分析表明,这些差异蛋白质主要定位于细胞外区,与肿瘤的生物合成、细胞增殖、血管生成相关。Western boltting和qRT-PCR验证了K1C19(下调0.11倍)和PSME2(上调2.02倍)的表达。结论:TMT定量蛋白组学方法能有效筛选有腋窝淋巴结转移的和无腋窝淋巴结转移的乳腺癌患者血清中的差异蛋白。其中,K1C19和PSME2是值得进一步研究的乳腺癌淋巴结转移的候选血清标志物。
OBJECTIVE: To analyze the differential proteins in sera of breast invasive ductal carcinoma patients with axillary lymph node metastasis and without axillary lymph node metastasis, and to screen biomarkers and therapeutic targets related to breast cancer metastasis. Methods: In this study, serum protein of 14 patients with axillary lymph node metastasis and 14 patients without axillary lymph node metastasis were detected by using tandem mass tag (TMT) labeling technique and the protein was quantified Analysis, screening out the significant changes in the differential proteins, and then search the Uniprot database and Proteome Discoverer software analysis for further bioinformatics analysis. Results: There were 119 differentially expressed proteins in axillary lymph node metastasis group, of which 6 were up-regulated and 113 down-regulated. Gene annotation and functional clustering analysis of gene ontology (GO) indicated that these differentially expressed proteins are mainly located in the extracellular domain, which is related to tumor biosynthesis, cell proliferation and angiogenesis. Western boltting and qRT-PCR validated the expression of K1C19 (down-regulated by 0.11-fold) and PSME2 (up-regulated by 2.02-fold). Conclusion: TMT quantitative proteomics method can effectively screen the differential proteins in the serum of patients with axillary lymph node metastasis and without axillary lymph node metastasis. Of these, K1C19 and PSME2 are candidate serum markers for further study of lymph node metastasis in breast cancer.