TXNDC5在胃癌细胞中相互作用蛋白分子的筛选  

Screening and identification of TXNDC5 binding protein in gastric cancer cells

作　　者：张林[1] 侯艳红[1] 李春梅[1] 张静[1] 雷迎峰 ZHANG Lin;HOU Yanhong;LI Chunmei;ZHANG Jing;LEI Yingfeng(Department of Gastroenterology,the 8th Medical Center of PLA General Hospital,Beijing 100091;Department of Microbiology,Air Force Military Medical University,China)

机构地区：[1]解放军总医院第八医学中心消化内科,北京100091 [2]解放军空军军医大学微生物学教研室

出　　处：《胃肠病学和肝病学杂志》2021年第2期140-144,共5页Chinese Journal of Gastroenterology and Hepatology

摘　　要：目的筛选并分析硫氧还蛋白5(TXNDC5)在胃癌细胞中的关键相互作用蛋白分子。方法以pcDNA3.1为基础构建pcDNA3.1-TXNDC5-FLAG真核表达载体,并以此载体瞬时转染人胃癌细胞系SGC7901,RT-PCR法及Western blotting法检测TXNDC5融合蛋白表达。采用串联亲和纯化技术收集胃癌细胞内TXNDC5相互作用蛋白分子进行电泳分离及ESI-Q-TOF串联质谱分析鉴定TXNDC5相互作用蛋白质。结果构建的pcDNA3.1-TXNDC5-FLAG真核表达载体经DNA序列测定完全正确,转染胃癌细胞后RT-PCR法及Western blotting法可检测到TXNDC5分子表达。经串联亲和纯化及蛋白质组学分析,通过NCBI蛋白质数据库检索比对,筛选出TXNDIP等16个蛋白分子。结论本研究通过串联亲和偶联蛋白质组学技术成功的筛选出胃癌细胞SGC7901中与TXNDC5相互作用的蛋白质,并进行了初步分析,提出了TXNDC5可能的分子作用机制,为进一步研究打下基础。Objective To screen and analyze the key interaction proteins of TXNDC5 in gastric cancer cells.Methods The eukaryotic expression vector pcDNA3.1-TXNDC5-FLAG was constructed based on pcDNA3.1 and transiently transfected into human gastric cancer cell line SGC7901.The expression of TXNDC5 fusion protein was detected by RT-PCR and Western blotting.TXNDC5 interacting proteins in gastric cancer cells were collected by tandem affinity purification technique,separated by electrophoresis and identified by ESI-Q-TOF tandem mass spectrometry.Results The eukaryotic expression vector pcDNA3.1-TXNDC5-FLAG was correctly sequenced.The expression of TXNDC5 was detected by RT-PCR and Western blotting after transfection.After tandem affinity purification and proteomic analysis,16 protein molecules such as TXNDIP were screened by searching NCBI protein database.Conclusion The proteins interacting with TXNDC5 in SGC7901 gastric cancer cells are successfully screened by tandem affinity coupled proteomics,and the possible molecular mechanism of TXNDC5 was proposed.

关 键 词：TXNDC5 相互作用蛋白 蛋白质组学 TXNIP 

分 类 号：R735.2[医药卫生—肿瘤]