基于TMT蛋白组学的硝酸铀酰诱导CHO-K1细胞损伤研究  

Study on injury of CHO-K1 cells induced by uranyl nitrate based on TMT proteomics

作　　者：尹晶晶[1] 刘欢[1] 高洁 王志鹏 袁慧[1] 李建华 李建国[1] YIN Jing-jing;LIU Huan;GAO Jie;WANG Zhi-peng;YUAN Hui;LI Jian-hua;LI Jian-guo(Department of Radiological and Environmental Medicine,Shanxi Key Laboratory of Drug Toxicology and Drug for Radiation Injury,CNNC Key Laboratory on Radiotoxicology and Radiopharmaceutical Preclinical Evaluation,China Institute for Radiation Protection,Taiyuan 030006,China)

机构地区：[1]中国辐射防护研究院放射医学与环境医学研究所药物毒理与放射损伤药物山西省重点实验室中核集团放射毒理与放射性药物临床前评价重点实验室,山西太原030006

出　　处：《陕西科技大学学报》2023年第1期66-71,87,共7页Journal of Shaanxi University of Science & Technology

基　　金：HCL科研专项基金项目(HC17110217)。

摘　　要：应用蛋白组学技术筛选硝酸铀酰暴露诱导CHO-K1细胞损伤的差异表达蛋白,为铀毒性机制研究提供数据.CHO-K1细胞经500μmol/L硝酸铀酰染毒24 h,以差异倍数大于1.5倍,差异显著性小于0.05为标准,采用串联质谱标签(tandem mass tag,TMT)标记蛋白组学技术筛选差异表达蛋白,应用生物信息学技术对差异表达蛋白进行聚类分析、基因本体(Gene Ontology,GO)分析、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析及蛋白相互作用分析.共鉴定出蛋白4772个,硝酸铀酰染毒组筛选出差异表达1.5倍以上的蛋白309个,其中164个表达上调,145个表达下调.GO分析结果显示差异表达的蛋白主要参与1086种生物过程、282种细胞成分以及377类分子功能.KEGG分析结果显示差异表达蛋白主要富集于40条信号转导通路,参与核糖体生物合成、RNA运输、辅因子代谢、Notch信号通路、吞噬体、染色体及相关蛋白、蛋白质外排、生理节律、IL-17信号通路、类脂物代谢等信号通路.差异蛋白相互作用网络分析结果显示蛋白RPS3A和RPS17的关联度最高.研究结果表明,CHO-K1细胞经500μmol/L硝酸铀酰染毒24 h后,蛋白发生差异表达,核糖体生物合成、RNA运输等多条信号通路发生改变,蛋白RPS3A和RPS17可能是硝酸铀酰暴露致CHO-K1细胞损伤的关键蛋白.Application of proteomics technology screening of uranyl nitrate exposure induced CHO-K1 differentially expressed proteins,provide data for the research of uranium toxicity mechanism.CHO-K1 cells were exposed to 500μmol/L uranyl nitrate for 24 h.Taking the difference factor more than 1.5 times and the difference significance less than 0.05 as the standard,tandem mass tag(TMT)proteomics technology was used to screen differentially expressed proteins,and bioinformatics technology was applied to differentially expressed proteins for cluster analysis,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis and protein interaction analysis.Protein identification results showed that 4772 proteins were identified.309 proteins with a differential expression of more than 1.5 times were screened in the uranyl nitrate exposure group,among which 164 proteins were up-regulated and 145 proteins were down-regulated.GO analysis results showed that the differentially expressed proteins were mainly involved in 1086 biological processes,282 cellular components and 377 molecular functions.KEGG analysis results showed that the differentially expressed proteins were mainly enriched in 40 signal transduction pathways.It is involved in ribosome biosynthesis,RNA transport,cofactor metabolism,Notch signaling pathway,phagosome,chromosome and related proteins,protein effection,circadian rhythm,IL-17 signaling pathway,lipid metabolism and other signaling pathways.Differential protein interaction network analysis showed that RPS3 A and RPS17 had the highest correlation.The results indicated that after exposure to 500μmol/L uranyl nitrate for 24 h,proteins were differentially expressed,ribosome biosynthesis,RNA transport and other signaling pathways were changed,and proteins RPS3 A and RPS17 may be the key proteins in the damage of CHO-K1 cells induced by uranyl nitrate exposure.

关 键 词：硝酸铀酰 CHO-K1细胞 蛋白质组学 生物信息学 差异蛋白 

分 类 号：R114[医药卫生—卫生毒理学]