CPB2为晚期肺腺癌EGFR-T790M耐药基因突变的潜在生物标志物  

CPB2 is a potential biomarker for EGFR-T790M resistance gene mutations in advanced lung adenocarcinoma

作　　者：王为民 韩志刚[1] 俞婷婷[1] Wang Weimin;Han Zhigang;Yu Tingting(Department of Pulmonary Medicine Ward 1,Affiliated Tumor Hospital of Xinjiang Medical University,Urumqi 830000,China)

机构地区：[1]新疆医科大学附属肿瘤医院肺内科一病区,新疆维吾尔自治区乌鲁木齐830000

出　　处：《实用肿瘤杂志》2024年第3期242-251,共10页Journal of Practical Oncology

基　　金：省部共建中亚高发病成因与防治国家重点实验室资助(SKL-HIDCA-2020-JZ)。

摘　　要：目的 通过平行反应监测(parallel reaction monitoring,PRM)定量蛋白组学技术验证晚期肺腺癌患者表皮生长因子受体(epidermal growth factor receptor,EGFR)-T790M耐药突变的生物标志物。方法 纳入2018年1月至2018年12月就诊于新疆医科大学附属肿瘤医院的口服EGFR-酪氨酸激酶抑制剂(tyrosine kinase inhibitor,TKI)吉非替尼的具有EGFR19del或EGFR 21 L858R基因突变的晚期肺腺癌患者44例。在患者疾病进展后行组织活检,使用下一代测序(next generation sequencing,NGS)方法检测出EGFR-T790M突变患者24例,非EGFR-T790M突变20例。提取44例患者的血清蛋白,通过SDS-PADE进行质量评估,两组分别得到10例质量合格血清蛋白样本。质量合格的血清蛋白样品分别进行丝氨酸蛋白酶酶解,通过PRM方法进行纳米液相色谱-串联质谱分析法(nanometer liquid chromatography-tandem mass spectrometry,nanoLC-MS/MS)检测。运行Skyline软件进行数据分析,得出与EGFR-T790M突变相关的差异表达蛋白。将基因本体论(Gene Ontology,GO)数据库的各节点映射,进行功能富集分析,并利用京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)数据库提供的绘图模块进行差异表达蛋白表达含量渲染,采用STRING数据库构建差异表达蛋白的蛋白互作(protein-protein interaction,PPI)网络,寻找关键差异表达蛋白。结果 通过PRM方法确定14种与晚期肺腺癌EGFRT790M突变相关的差异表达蛋白。GO富集分析显示,这些差异表达蛋白在参与细胞组分方面,主要构成细胞外区域的部分;在发挥的分子功能方面,主要为抗原结合、作用于L氨基酸肽的肽酶活性和免疫球蛋白受体结合。KEGG富集通路分析显示,补体和凝血级联反应差异表达蛋白富集指数最高。该通路上的差异蛋白为凝血因子Ⅹ(coagulation factorⅩ,F10)、F9、纤溶酶原(plasminogen,PLG)和羧肽酶B2(carboxypeptidase B2,CPB2)。其中F10为上调蛋白,其余为Objective To verify the presence of epidermal growth factor receptor(EGFR)-T790M resistance mutant biomarkers in patients with advanced lung adenocarcinoma using the quantitative proteomic approach known as parallel reaction monitoring(PRM).Methods Forty-four advanced lung adenocarcinoma patients with EGFR 19del or EGFR 21 L858R mutation and treated with EGFR-ty-rosine kinase inhibitor(TKI)gefitinib at the Affiliated Tumor Hospital of Xinjiang Medical University from January 2018 to December 2018 were included in this study.Tissue biopsies were conducted after disease progression.Next generation sequencing(NGS)was employed to identify EGFR-T790M mutation in 24 patients and no EGFR-T790M mutation in 20 patients.Forty-four patients'serum proteins were ex-tracted and evaluated using SDS-PAGE.High-quality serum protein samples were obtained from 10 patients with EGFR-T790M mutation and 10 without EGFR-T790M mutation.These high-quality samples were individually digested with serine protease enzymes and then an-alyzed using nanometer liquid chromatography-tandem mass spectrometry(nanoLC-MS/MS)via the PRM method.The resulting data were analyzed using Skyline software to identify the differentially expressed proteins associated with EGFR-T790M mutation.The genes were assigned to the nodes of the Gene Ontology(GO)database,and functional enrichment analysis was conducted.The expression patterns of the differentially expressed proteins were visualized using the mapping module provided by the Kyoto Encyclopaedia of Genes and Genomes(KEGG)database.The STRING database was utilized to construct protein-protein interaction(PPI)networks of the differentially expressed proteins in order to identify key differentially expressed proteins.Results The PRM technique successfully identified 14 distinct proteins that were differently expressed in advanced lung adenocarcinoma with EGFR-T790M mutation.The GO enrichment analysis revealed that the 14 differentially expressed proteins primarily belonged to the extracellular region in terms of

关 键 词：晚期肺腺癌 EGFR-T790M 蛋白组学 差异性蛋白 

分 类 号：R734.2[医药卫生—肿瘤]