pEGFP-N1-IRF3a重组质粒通过调节STAT1的活化诱导非小细胞肺癌细胞的凋亡  

Recombinant pEGFP-N1-IRF3a Plasmid Induces the Apoptosis of Non-Small Cell Lung Cancer Cells by Regulating the Activity of STAT1

作　　者：易亮 吴艳萍 韩倩[2] 杨云梅[1] YI Liang;WU Yanping;HAN Qian;YANG Yunmei(Department of Geriaric Medicine,the First Afiliated Hospital,College of Medicine,Zhejiang University,Hangzhou 310003,China;Department of Geriatrics,Peking University First Hospital,Bejing 10034,China)

机构地区：[1]浙江大学医学院附属第一医院老年医学科,杭州310003 [2]北京大学第一医院老年内科,北京100034

出　　处：《中国细胞生物学学报》2020年第3期385-391,共7页Chinese Journal of Cell Biology

摘　　要：干扰素调节因子3(interferon regulatory factor 3,IRF3)在固有免疫激活的过程中发挥重要作用,其中IRF3a是由IRF3可变剪接形成的一个亚型,目前几乎没有研究报道该蛋白对肿瘤细胞凋亡的影响。在该研究中,首先通过qRT-PCR检测IRF3a基因在肺癌组织以及癌旁组织中的表达水平;然后通过PCR从人外周血细胞中获取IRF3a基因,与质粒pEGFP-N1成功构建重组质粒pEGFP-N1-IRF3a。重组质粒转染非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞A549、H1299后,细胞凋亡比例明显升高,cleaved caspase8、cleaved caspase3水平也显著升高。此外,过表达IRF3a能激活STAT1,以p-STAT1抑制剂Nifuroxazide抑制STAT1的活性能显著抑制IRF3a诱导的细胞凋亡。因此,该研究成功构建了pEGFP-N1-IRF3a重组质粒,并进一步证明了IRF3a通过激活STAT1诱导NSCLC细胞的凋亡。IRF3 (interferon regulatory factor 3) plays the significant roles in regulating innate immune activity.IRF3a (interferon regulatory factor 3 isoform 3) is the translated production of IRF3 transcript variant 3 under the control of alternative splicing.However,seldom researches whether IRF3a has effect on the apoptosis of cancer cells been performed.In this research,the level of IRF3a gene expression was firstly tested in the lung cancer tissues and the adjacent tissues by qRT-PCR.IRF3a gene was obtained from human peripheral blood mononuclear cells by PCR and cloned into pEGFP-N1 plasmid,then the recombinant pEGFP-N1-IRF3a plasmid was successfully constructed.The apoptotic cells dramatically increased and the level of cleaved caspase3,cleaved caspase8 strikingly elevated followed recombinant pEGFP-N1-IRF3a plasmid transfected into A549 and H1299 cells.Moreover,IRF3a overexpression could promote the activity of STAT1 and the apoptotic cells decreased obviously following the pretreatment of STAT1 inhibitor Nifuroxazide.Therefore,recombinant pEGFP-N1-IRF3a plasmid was successfully constructed in this research and it further revealed that IRF3a promoted the apoptosis of NSCLC A549 and H1299 cells by activating the phosphorylation of STAT1.

关 键 词：干扰素调节因子3a 质粒构建 非小细胞肺癌细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]