分次低剂量辐射诱导HBE细胞衰老中mRNA差异表达分析  

Differential mRNA expression in fractional low-dose radiation-induced senescence of HBE cells

作　　者：张灵钰 黄伟旭[2] 蔡雅诗 李慧娴 张敏 闻昌勇 杨萍 邹剑明[2] 陈慧峰 ZHANG Lingyu;HUANG Weixu;CAI Yashi;LI Huixian;ZHANG Min;WEN Changyong;YANG Ping;ZOU Jianming;CHEN Huifeng(School of Public Health,Guangzhou Medical University,Guangzhou 511436 China;Guangdong Province Hospital for Occupational Disease Prevention and Treatment,Guangzhou 510300 China;School of Public Health,Southern Medical University,Guangzhou 510515 China)

机构地区：[1]广州医科大学公共卫生学院,广东广州511436 [2]广东省职业病防治院,广东广州510300 [3]南方医科大学公共卫生学院,广东广州510515

出　　处：《中国辐射卫生》2024年第2期116-122,共7页Chinese Journal of Radiological Health

基　　金：国家自然科学基金(81302387);广东省基础与应用基础研究基金(2019A1515011969、2022A1515012421、2023A1515010414);广州市科技计划项目(202002030031)。

摘　　要：目的利用高通量mRNA测序和生物信息学技术探讨分次低剂量电离辐射(low dose ionizing radiation,LDIR)诱导人支气管上皮(human bonchial epithelial,HBE)细胞衰老中差异表达mRNA及相关生物学过程与通路。方法分别采用0、50、100、200 mGy剂量辐照HBE细胞7次结束后24和48 h收集细胞,检测衰老相关β-半乳糖苷酶活性和衰老相关分泌表型基因mRNA和蛋白表达水平,采用高通量测序筛选差异表达基因进行基因本体学(GO)和京都基因与基因组大百科全书(KEGG)分析。结果分次低剂量辐照HBE细胞衰老阳性面积呈剂量依赖性增加(P<0.05)。与对照组相比,辐照结束后24和48 h,100 mGy×7和200 mGy×7组转化生长因子β1(transforming growth factor-β1,TGF-β1)与基质金属蛋白酶9(matrix metalloprotein-9,MMP-9)基因mRNA水平与蛋白表达均增加。高通量测序显示,与对照组相比各剂量组差异表达mRNA分别有882个、475个和1205个。GO分析表明,各剂量组差异表达mRNA主要富集在细胞周期调节、氮化合物代谢过程的调控、对刺激反应的调节和细胞分裂等生物过程。KEGG分析显示,差异表达mRNA主要富集在细胞周期、细胞衰老、铁死亡等通路。结论分次LDIR可诱导HBE细胞衰老,衰老细胞中差异表达mRNA相关生物学过程和通路与细胞周期、细胞衰老等有关。Objective To explore the differentially expressed mRNAs and related biological processes and pathways in fractional low-dose ionizing radiation(LDIR)-induced senescence of normal human bronchial epithelial(HBE)cells by highthroughput mRNA sequencing and bioinformatics techniques.Methods Senescence-associatedβ-galactosidase staining and senescence-associated secretion phenotype gene mRNA and protein expression levels were measured at 24 and 48 h after irradiating HBE cells 7 times at doses of 0,50,100,and 200 mGy,respectively.The differentially expressed genes were screened by high-throughput sequencing for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses.Results The senescence-positive area of fractional low-dose irradiated HBE cells increased in a dose-dependent manner(P<0.05).The mRNA levels and protein expression of transforming growth factor-β1(TGF-β1)and matrix metalloproteinase-9(MMP-9)genes were increased in the 100 mGy×7 and 200 mGy×7 groups at 24 and 48 h after the end of irradiation compared with the control group.High-throughput sequencing showed that there were 882,475,and 1205 differentially expressed mRNAs in each dose group compared with the control group.GO analysis showed that the differentially expressed mRNAs in each dose group were mainly enriched in biological processes such as cell cycle regulation,regulation of nitrogen compound metabolic process,regulation of cell division and response to stimulus.KEGG analysis showed that the differentially expressed mRNAs were mainly enriched in the pathways of cell cycle,cell senescence,and ferroptosis.Conclusion Fractional LDIR induced senescence in HBE cells,and differentially expressed mRNA-associated biological processes and pathways in senescent cells are related to cell cycle and cell senescence.

关 键 词：低剂量电离辐射 HBE细胞 衰老 

分 类 号：Q691[生物学—生物物理学]