醛脱氢酶18家族成员A1在食管癌细胞中结合的靶标及测序分析
Binding target and sequencing analysis of aldehyde dehydrogenase 18 family member A1 in esophageal cancer cells
王永康 1余玮 1克力木·阿不都热依木 2买买提·依斯热依力3
作者信息
- 1. 新疆医科大学研究生学院(乌鲁木齐 830017)
- 2. 新疆维吾尔自治区人民医院自治区普外微创研究所(乌鲁木齐 830001)
- 3. 新疆医科大学研究生学院(乌鲁木齐 830017);新疆维吾尔自治区人民医院自治区普外微创研究所(乌鲁木齐 830001);新疆维吾尔自治区人民医院科研教育中心(乌鲁木齐 830001)
- 折叠
摘要
目的 研究醛脱氢酶 18 家族成员 A1(aldehyde dehydrogenase 18 family member A1,ALDH18A1)作为一种RNA结合蛋白在食管癌细胞(KYSE150细胞)中结合RNA的分子特征及对肿瘤生长的影响.方法 通过体外培养人食管鳞癌细胞(KYSE150细胞).同时,采用RNA免疫共沉淀技术研究细胞内RNA与蛋白结合情况,利用目标蛋白的抗体将相应的RNA-蛋白复合物沉淀下来,分离纯化捕获的RNA.分析ALDH18A1结合RNA的分子特征,并且对ALDH18A1结合靶基因进行京都基因与基因组百科全书聚类分析.结果 蛋白质免疫印迹实验显示目的蛋白被抗体较好富集.ALDH18A1具有广泛的RNA结合活性,结合显著富集在编码区、内含子和5'非翻译区.ALDH18A1主要结合在RNA的UGUAAUC基序.聚类分析结果表明,ALDH18A1结合的RNA分子主要参与了在对焦点黏附、癌症的中心碳代谢、细胞周期、剪接体、RNA运输和泛素介导的蛋白水解等.结论 ALDH18A1能结合RNA分子,可在食管癌相关基因的表达过程以及相关的生物学过程过程中发挥作用.
Abstract
Objective To study the molecular characteristics of RNA binding protein aldehyde dehydrogenase 18 family member A1(ALDH18A1)in esophageal carcinoma cells(KYSE150 cells)and its effect on tumor growth.Methods Human esophageal squamous cell(KYSE150 cells)was cultured in vitro.At the same time,RNA co-immuno precipitation technology was used to study the binding of RNA and protein in the cell,and the corresponding RNA-protein complex was precipitated by the antibody of the target protein to separate and purify the captured RNA.The molecular characteristics of ALDH18A1 binding RNA were analyzed,and KyotoEncyclopedia of Genes and Genomes cluster analysis was performed for ALDH18A1 binding target genes.Results Protein immunoblotting experiments showed that the target protein was well enriched by antibodies.ALDH18A1 had extensive RNA binding activity,with significant enrichment in regions such as coding sequences,intron,and 5'untranslated region.ALDH18A1 mainly bound to the UGUAAUC motif of RNA.The cluster analysis showed that the RNA molecules bound to ALDH18A1 mainly participated in focal adhesion,central carbon metabolism in cancer,cell cycle,spliceosome,RNA transport,and ubiquitin mediated protein hydrolysis.Conclusion ALDH18A1 has the function of binding to RNA molecules and may play a role in the expression of esophageal cancer-related genes and related biological processes.
关键词
食管癌细胞/醛脱氢酶18家族成员A1/RNA结合蛋白/RNA免疫共沉淀Key words
Esophageal cancer cells/aldehyde dehydrogenase 18 family member A1/RNA binding protein/RNA immunocoprecipitation引用本文复制引用
基金项目
新疆医科大学研究生创新创业训练计划(CXCY2024017)
新疆维吾尔自治区"天山英才"培养计划(2023TSYCCX0056)
新疆维吾尔自治区优秀博士后普通基金(346798)
出版年
2024
NETL
NSTL
万方数据