摘要
目的探讨miR-6129在二氢二醇环氧苯并芘(BPDE)致BEAS-2B细胞DNA损伤中的作用。方法以人正常肺上皮细胞BEAS-2B作为对照,采用实时荧光定量聚合酶链反应(RT-qPCR)检测在不同剂量BPDE暴露下,BEAS-2B细胞中miR-6129的表达水平变化;采用CCK-8法分别检测过表达miR-6129和干扰miR-6129表达对BPDE暴露下细胞活性影响;通过单细胞凝胶电泳实验(彗星实验)分析miR-6129高表达和miR-6129低表达对BPDE致细胞DNA损伤的影响。结果不同剂量的BPDE染毒可诱导人正常肺上皮BEAS-2B细胞miR-6129表达水平的升高,且呈剂量—反应关系趋势(P<0.05);在BPDE染毒后,转染miR-6129 mimic的miR-6129高表达组细胞的存活率均明显下降,差异有统计学意义(P<0.05),同时彗星实验显示DNA损伤程度增加;而miR-6129低表达组细胞在BPDE染毒后细胞存活率明显增加(P<0.05),DNA损伤程度降低(P<0.05)。结论BPDE可能通过上调BEAS-2B细胞中miR-6129表达,导致细胞DNA损伤。
Objective To explore the role of miR-6129 in DNA damage of human normal lung epithelial BEAS-2B cells induced by BPDE exposure.Methods Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of miR-6129 in BEAS-2B cells exposed to different doses of BPDE.The CCK-8 method was used to detect the effects of overexpression of miR-6129 and interference with miR-6129 expression on cell viability exposed to BPDE.The comet assay was used to analyze the effects of overexpression of miR-6129 and interference with miR-6129 expression on cell DNA damage exposed to BPDE.Results The expression levels of miR-6129 were significantly increased induced by BPDE treatment with a dose-response relationship,compared with control cells(P<0.05).After BPDE exposure,the survival rate of miR-6129 high expression group was significantly decreased(P<0.05),and the comet assay showed that the degree of DNA damage was increased.However,the cell viability of miR-6129 low expression group was higher than that of control group(P<0.05),and the degree of DNA damage was decreased(P<0.05).Conclusion BPDE may cause cellular DNA damage by upregulating miR-6129 expression in BEAS-2B cells.
作者
李梦承
张诚
梁琳琳
刘嘉禹
周家圳
杨巧媛
LI Meng-cheng;ZHANG Cheng;LIANG Lin-lin;LIU Jia-yu;ZHOU Jia-zhen;YANG Qiao-yuan(Institute of Chemical Carcinogenesis,Guangzhou Medical University,Guangzhou Guangdong 511436,China)
出处
《毒理学杂志》
CAS
CSCD
2022年第4期280-285,291,共7页
Journal of Toxicology
基金
国家自然科学基金(81773385)
广东省省级科技计划项目(2017A020215066)
广东省基础与应用基础研究基金项目(2019A1515011298,2022A1515010727)。
