矽肺患者周围血单个核细胞中差异表达miRNA筛选与验证  

Screening and verification of differentially expressed miRNAs in PBMCs of patients with silicosis

作　　者：赵英政[1,2] 常美玉 李海斌 关毅[2] 李宁 白义春 李玉春 徐光翠 姚三巧[1,2] ZHAO Ying-zheng;CHANG Mei-yu;LI Hai-bin;GUAN Yi;LI Ning;BAI Yi-chun;LI Yu-chun;XU Guang-cui;YAO San-qiao(School of Public Health,Xinxiang Medical University,Xinxiang,Henan 453003,China;不详)

机构地区：[1]新乡医学院公共卫生学院,河南新乡453003 [2]华北理工大学公共卫生学院,河北唐山063000

出　　处：《中国职业医学》2022年第3期248-253,共6页China Occupational Medicine

基　　金：国家自然科学基金项目(81773399);中国煤炭工业协会2018年指导性计划项目(MTKJ-2018284);华北理工大学研究生创新项目(2017B12)。

摘　　要：目的筛选和验证矽肺患者周围血单个核细胞(PBMCs)中差异表达miRNAs。方法采用随机数表法,选择48例职业性矽肺壹期患者(病例组)和45名健康对照人员(对照组)为研究对象,各采集周围血以Ficoll-Paque梯度离心法分离PBMCs。从2组人员中各随机选择3人进行miRNAs转录组测序,采用R Studio软件筛选差异表达miRNAs,采用FunRich软件预测PBMCs中差异表达miRNAs相关的上游转录因子,采用实时荧光定量聚合酶链反应验证其余样本的差异miRNAs的表达水平。结果共筛选出124个差异表达的miRNAs,其中97个miRNAs表达上调,27个miRNAs表达下调。根据差异miRNAs预测获得的67个靶基因主要参与了细胞内过程和核酸结合转录因子活性等分子过程,包括细胞粘附分子、大鼠肉瘤、破骨细胞分化等通路。差异表达上调的前5个miRNAs为hsa-miR-137-3p、hsa-miR-500b-3p、hsa-miR-190a-5p、hsa-miR-141-3p和hsa-miR-22-3p,差异表达下调的前5个miRNAs为hsa-miR-202-5p、hsa-miR-548ai、hsa-miR-5587-3p、hsa-miR-570-5p和hsa-miR-10398-3p;差异表达变化趋势与测序结果一致。PBMCs中差异表达miRNAs相关的上游转录因子包括特异性蛋白1、早期生长反应蛋白1和锌指蛋白161等。结论矽肺患者PBMCs中差异表达miRNAs及其分子通路可能在二氧化硅粉尘致PBMCs损伤中发挥调控作用。ObjectiveTo screen and validate the differentially expressed micro RNAs(mi RNAs)in peripheral blood mononuclear cells(PBMCs)of patients with silicosis.MethodsForty-eight patients with occupational silicosis at stageⅠ(case group)and 45 healthy controls(control group)were selected as research subjects by random number table method.PBMCs were separated by Ficoll-Paque gradient centrifugation from peripheral blood.Three people from each group were randomly selected for mi RNAs transcriptome sequencing.R Studio software was used to screen differentially expressed mi RNAs,and Fun Rich software was used to predict the upstream transcription factors related to the differentially expressed mi RNAs in PBMCs.The differentially expressed mi RNAs were verified by real-time quantitative polymerase chain reaction.ResultsA total of 124differentially expressed mi RNAs were screened,among them,97 mi RNAs were up-regulated and 27 mi RNAs were down-regulated.The 67 target genes predicted by differential mi RNAs were mainly involved in intracellular processes and nucleic acid-binding transcription factor activities,including cell adhesion molecules,rat sarcoma,osteoclast differentiation and other pathways.The top five mi RNAs differentially up-regulated were hsa-mi R-137-3p,hsa-mi R-500b-3p,hsa-mi R-190a-5p,hsa-mi R-141-3p and hsa-mi R-22-3p.The top five differentially down-regulated mi RNAs were hsa-mi R-202-5p,hsa-mi R-548ai,hsa-mi R-5587-3p,hsa-mi R-570-5p and hsa-mi R-10398-3p.The change of these mi RNAs were consistent with the results predicted according to the mi RNAs transcriptome sequencing.The transcription factors including specificity protein 1,early growth response 1,zinc finger protein 161,etc.,were obtained according to the differentially expressed mi RNAs.ConclusionThe differentially expressed mi RNAs and corresponding molecular pathways play a role in the regulation of PBMCs cytotoxicity induced by silica particles.

关 键 词：矽肺 周围血 PBMCS 差异表达 MIRNAS 转录因子 筛选 验证 

分 类 号：R135.2[医药卫生—劳动卫生]