摘要
目的:基于微小RNA 146a(miR-146a)和肿瘤坏死因子受体相关蛋白6(TRAF6)探究长链非编码RNA(LncRNA)NORAD对非霍奇金淋巴瘤(NHL)细胞焦亡、增殖和转移的影响。方法:培养NHL细胞系Jurkat、Raji和正常B细胞,通过实时荧光定量聚合酶链式反应(qRT-PCR)检测LncRNA NORAD、miR-146a和TRAF6表达水平。选择Raji细胞系进行LncRNA NORAD表达敲低处理(NORAD-siNC组,NORAD-si组),通过Western blot检测NOD样受体蛋白3(NLRP3)、Cleaved caspase1、Gasdermins-DN端体(N-GSDMD)、TRAF6表达水平变化,qRT-PCR检测miR-146a表达水平变化,试剂盒检测细胞培养上清中白介素-1β(IL-1β)、IL-18水平变化,EDU实验检测细胞增殖活性变化,Transwell实验检测细胞侵袭活性变化。之后,通过检测双萤光素酶活性验证LncRNA NORAD和miR-146a、miR-146a和TRAF6之间的相关性。结果:与正常B细胞相比,LncRNA NORAD和TRAF6在Jurkat、Raji细胞中表达升高,miR-146a在Jurkat、Raji细胞中表达降低。与NORAD-siNC组细胞相比,NORAD-si组细胞miR-146a、NLRP3、N-GSDMD、Cleaved caspase1表达水平升高,TRAF6表达水平降低,细胞增殖和侵袭活性减弱,细胞培养上清中IL-1β和IL-18水平均升高。双萤光素酶检测结果表明,LncRNA NORAD和miR-146a、miR-146a和TRAF6之间均存在靶向结合关系。结论:LncRNA NORAD在NHL细胞中表达上调,敲低LncRNA NORAD表达可以通过miR-146a/TRAF6轴诱导NHL细胞焦亡,抑制细胞增殖和侵袭。
Objective:To investigate the effect of lncRNA NORAD on pyroptosis,proliferation and invasion of non-Hodgkin's lymphoma cells based on miR-146a/TRAF6 axis.Methods:NHL cell lines Jurkat,Raji and normal B cells were cultured.The expression levels of lncRNA NORAD,miR-146a and TRAF6 were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Raji cell line was selected for lncRNA NORAD knockdown treatment(NORAD-siNC group,NORAD-si group).Western blot was used to detect the expression levels of NOD-like receptor protein 3(NLRP3),Cleaved caspase1,Gasdermins-DN terminal domain(N-GSDMD)and TRAF6.qRT-PCR was used to detect the expression level of miR-146a.The levels of interleukin-1β(IL-1β)and IL-18 in cell culture supernatant were detected by kits.EDU assay was used to detect cell proliferation activity,and Transwell assay was used to detect cell invasion activity.Then,the correlation between lncRNA NORAD and miR-146a,and between miR-146a and TRAF6 was verified by dual-luciferase activity detection.Results:Compared with normal B cells,the expression of lncRNA NORAD and TRAF6 was increased in Jurkat and Raji cells,while the expression of miR-146a was decreased.Compared with cells in the NORAD-siNC group,cells in the NORAD-si group had increased expression levels of miR-146a,NLRP3,N-GSDMD and Cleaved caspase1,decreased expression level of TRAF6,weakened cell proliferation and invasion activities,and increased levels of IL-1βand IL-18 in cell culture supernatant.The results of dual-luciferase assay showed that there were target binding relationships between lncRNA NORAD and miR-146a,and between miR-146a and TRAF6.Conclusion:lncRNA NORAD is up-regulated in NHL cells.Knocking down the expression of lncRNA NORAD can induce pyroptosis of NHL cells and inhibit cell proliferation and invasion through the miR-146a/TRAF6 axis.
作者
李杨
刁玉巧
王江娅
朱秀丽
江莲
LI Yang;DIAO Yuqiao(The Fourth Hospital of Hebei Medical University,Hebei Shijiazhuang 050000,China)
出处
《河北医学》
2025年第7期1087-1092,共6页
Hebei Medicine
基金
2023年度河北省医学科学研究课题计划,(编号:20230915)。
