基于RNA-seq技术分析模拟微重力的作用机制  

Mechanism of simulated microgravity based on RNA-seq technology

作　　者：秦玲玫 金蕊 周影 李江波 李心悦 程龙 QIN Lingmei;JIN Rui;ZHOU Ying;LI Jiangbo;LI Xinyue;CHENG Long(Institute of Biotechnology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;The 928th Hospital of the Joint Logistic Support Force of Chinese PLA,Haikou,Hainan 570100,China;Bioinformatics Center,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]军事科学院军事医学研究院生物工程研究所,北京100850 [2]解放军联勤保障部队第九二八医院,海口570100 [3]军事科学院军事医学研究院生物信息中心,北京100850

出　　处：《军事医学》2023年第7期481-486,共6页Military Medical Sciences

基　　金：科技部重点研发计划(2022YFC3600100);国家自然科学基金项目(82072717)。

摘　　要：目的 利用RNA转录组测序技术(RNA-seq)筛选模拟微重力(SMG)作用前后的差异表达基因(DEG),并探讨SMG在转录组学层面对人体细胞的作用机制。方法 按照不同培养条件将人白血病细胞HL-60分为3组:常规重力(NG)组、SMG组和SMG-NG(SMGNG)组,利用RNA-seq筛选SMG作用前后HL-60细胞mRNA DEG;用基因本体(GO)和京都基因与基因组百科全书(KEGG)进行功能注释,分析SMG调节的通路及在恢复NG后可快速回复及不可快速回复的通路。结果 与NG组相比,SMG组中共检测到735个DEG(424个上调和311个下调),其中有624个DEG的表达在SMGNG中可快速回复,110个DEG的表达在SMGNG中不可快速回复;GO、KEGG分析发现可快速回复的DEG与信号转导、脂质代谢、免疫调节及细胞凋亡高度相关,不可快速回复的DEG主要参与核苷酸结合寡聚化结构域(NOD)样受体信号转导相关通路、类固醇代谢和骨代谢过程。结论 初步明确了SMG调控的靶基因及信号通路,为进一步研究微重力的生物学功能及机制奠定了基础。Objective To screen differentially expressed genes(DEGs)before and after simulated microgravity(SMG)using RNA sequencing(RNA-seq),and to explore the mechanism of SMG for human cells at the transcriptomic level.Methods HL-60 cells were cultured under different culture conditions,the experiments were divided into three groups:the normal gravity(NG)group,simulated microgravity(SMG)group and SMG-NG(SMGNG)group.RNA-seq analysis was used to screen the differential expression profiles of mRNA in HL-60 cells before and after SMG.Gene ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were used for functional annotation to analyze the pathways regulated by SMG and those that could or could not recover rapidly after resumption of NG conditions.Results A total of 735 differentially expressed genes(DEGs)(424 up-regulated and 311 down-regulated)were identified in SMG comparing with NG,624 of which rapidly recovered in SMGNG,while 110 DEGs failed to rapidly recover.GO and KEGG analysis found that DEGs that rapidly recovered were highly correlated with signal transduction,lipid metabolism,immune regulation and apoptosis,while those that did not rapidly recover were mainly involved in pathways related to nucleotide-binding oligomerization domain(NOD)-like receptor signal transduction,steroid metabolism and bone metabolism.Conclusion The target genes and signaling pathways regulated by SMG are identified,which will facilitate research on the biological functions and mechanisms of microgravity.

关 键 词：模拟微重力 HL-60细胞 RNA-SEQ 转录组 差异表达基因 信号通路 

分 类 号：Q753[生物学—分子生物学] R852.22[医药卫生—航空、航天与航海医学]