绝经后骨质疏松症关联长链非编码RNA uc431+相互作用蛋白质组学分析  

Interaction proteomics of long noncoding RNA uc431+gene in postmenopausal osteoporosis

作　　者：李生强 谢冰颖 陈娟 谢丽华 黄景文 葛继荣 Li Shengqiang;Xie Bingying;Chen Juan;Xie Lihua;Huang Jingwen;Ge Jirong(Department of Osteoporosis Syndrome Genomics,Institute of Basic Research,Fujian Academy of Chinese Medical Sciences,Fuzhou 350003,Fujian Province,China)

机构地区：[1]福建省中医药科学院基础研究所骨质疏松证候基因组学研究室,福建省福州市350003

出　　处：《中国组织工程研究》2021年第11期1641-1646,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81774350),项目负责人:李生强;福建省自然科学基金项目(2019J01339),项目负责人:谢冰颖。

摘　　要：背景:过表达LncRNA TUG1促进破骨细胞的增殖及抑制凋亡,采用siRNA敲低则取得了相反的结果,表明敲低LncRNA TUG1可能成为一个骨质疏松治疗的潜在靶点。目的:研究与绝经后骨质疏松症关联非编码RNA uc431+相互作用的蛋白组并进行生物信息学分析。方法:将甲醛交联的人单核细胞白血病细胞株(THP-1)细胞超声破碎,实验组与结合生物素标记的特异性探针的磁珠进行杂交,对照组与非特异性探针的磁珠进行杂交,获得的与非编码RNA uc431+结合的多肽进行质谱分析,获得的数据使用MaxQuant进行搜库和定量分析。对2组样品的定量结果进行统计学分析,得到对应的富集蛋白,并进行GO、KEGG通路、蛋白互作分析和展示,构建蛋白质相互作用网络。结果与结论:①获得的多肽总数为918条,蛋白总数为271个,过滤后蛋白总数为241个,与对照组相比,实验组差异蛋白为10个:DDOST、DMBT1、HPD、IGLL5、IGK、LTF、LYZ、MUC5AC、PIGR及RPL23;②GO富集分析表明,它们参与防御反应、白细胞激活等生物学过程及核糖体rRNA结合、溶菌酶活性等分子功能;③KEGG通路分析预测它们参与辅酶Q生物合成、苯丙氨酸代谢、核糖体信号肽及唾液分泌等信号通路;④结合蛋白质组学及生物信息学分析,预测绝经后骨质疏松症关联非编码RNA uc431+可能通过互作蛋白参与免疫调节及骨代谢过程。BACKGROUND:Overexpression of LncRNA TUG1 promotes osteoclast proliferation and inhibits apoptosis.Knockdown with siRNA has achieved the opposite result,indicating that knockdown of LncRNA TUG1 may become a potential target for osteoporosis treatment.OBJECTIVE:To study the proteins interacting with the non-coding RNA uc431+associated with postmenopausal osteoporosis and carry out bioinformatics analysis.METHODS:Human monocytic leukemia cell line(THP-1)cells crosslinked with formaldehyde were broken by ultrasound.The experimental group was hybridized with magnetic beads combined with biotin labeled specific probes.The control group was hybridized with the magnetic beads of non-specific probes.The obtained peptides were identified using mass spectrometry.The data were searched and quantified by MaxQuant.The quantitative results of the two sets of samples were statistically analyzed,and the corresponding enrichment proteins were obtained.Gene ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,protein-protein interaction analysis and display were performed,and the protein-protein interaction network was constructed.RESULTS AND CONCLUSION:The total number of peptides obtained was 918,with 271 proteins in total,and the total number of proteins after filtration was 241.Compared with the control group,there were 10 differential proteins in the experimental group,including DDOST,DMBT1,HPD,IGLL5,IGK,LTF,LYZ,MUC5AC,PIGR,and RPL23.GO enrichment analysis showed that they were involved in biological processes such as defense reaction,leukocyte activation,ribosomal rRNA binding,lysozyme activity and other molecular functions.KEGG pathway analysis predicted that they were involved in ubiquinone and other terpenoid-quinone biosynthesis,phenylalanine metabolism,ribosome and salivary secretion.Combined with the analysis of proteomics and bioinformatics,it is predicted that uc431+,a gene related to postmenopausal osteoporosis,may be involved in immune regulation and bone metabolism by interacting proteins.

关 键 词：骨质疏松症 绝经 非编码RNA 蛋白组学 质谱 乳铁蛋白 骨代谢 生物信息学 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]