本文主要研究内容
作者王棋（2019）在《IFNγR2的转录调控机制研究》一文中研究指出：干扰素γ(interferon-γ,IFN-γ)在先天免疫、炎症反应、适应性免疫、肿瘤免疫等生命过程中发挥至关重要的作用。IFN-γ结合膜表面受体IFN-γR并激活其下游的JAK1、JAK2,进一步磷酸化转录因子STAT1,STAT1进入细胞核内结合靶基因.的特征性GAS(Gamma interferon activation site)元件,启动干扰素刺激基因(interferon stimulated gene,ISG)的表达。ISG中包含有趋化因子、抗原呈递分子、吞噬受体和各种具.有抗.菌抗病毒效应的细胞因子,参与细胞内病原体的防御,炎症与免疫反应调节,肿瘤免疫等免疫过程。IFN-γR由两种亚基组成:IFNγR1和IFNγR2。IFNγR1表达相对过剩,IFN-γ信号通路反应性水平由IFNγR2的表达量决定。因此阐明IFNγR2的调控机制有助于深入理解与IFN-γ信号通路有关的免疫反应和疾病发生过程,具有潜在的临床意义。目前关于IFNγR2的研究主要涉及其定位及功能,其转录调控机制研究少见报道。本研究拟通过构建IFNγR2荧光报告系统,为发现可调控IFNγR2转录水平的基因、小分子药物,进一步探索其调控方式和及其分子机制等提供有力工具。我们将小鼠Ifngr2基因的启动子序列放置于绿色荧光蛋白开放阅读框架上游,并将其共同插入慢病毒载体中,进一步通过慢病毒感染系统构建稳定表达Ifngr2启动子调控EGFP表达的Myc-Cap细胞系,以此细胞系用作研究IFNγR2调控的荧光报告体系。我们发现肿瘤坏死因子(tumor necrosis factor alpha,TNF-α)和奥沙利铂:(oxaliplatin):可以明显:上调Ifngr2基因的转录水平。TNF-α是机体系统性炎症急性期的重要细胞因子,主要由激活的巨噬细胞分泌,调控各种免疫过程并可诱导肿瘤细胞凋亡。TNF-α结合细胞表面的TNFR,激活下游NF-κB信号通路和MAPK信号通路。我们发现过表达NF-κB的亚基p65可上调Ifngr2。分别用IKK-β抑制剂IKK2 inhibitor IV、ML120B和蛋白酶体抑制剂MG132抑制NF-κB信号通路时候的激活,TNF-α均不能上调Ifngr2。这表明TNF-α通过NF-κB信号通路调控Ifngr2,这种调控发生在转录水平。MAPK信号通路抑制剂U1026、SP100625不能阻断TNF-α促进Ifngr2上调,提示MAPK信号通路可能不参与TNF-α上调Ifngr2的过程。为进一步明确Ifngr2启动子中关键的转录元件,我们构建小鼠Ifngr2基因启动子荧光素酶报告系统,通过系列截短分析探索Ifngr2基因启动子中转录调控的关键区域。生物信息学工具分析发现Ifngr2基因启动子-111--102片段GGGAAAGTCC为NF-κB结合位点,并通过突变分析对其进行了确认。Oxaliplatin作为一种抗肿瘤药物,广泛应用于多种肿瘤的化疗方案中。在细胞水平我们发现oxaliplatin.上调IFNγR2。我们建立了Myc-Cap皮下移植瘤动物模型型在动物水平初步探索了oxaliplatin对IFNγR2的调控作用和抗肿瘤免疫效应。本研究中我们构建了IFNγR2荧光报告体系,发现了TNF-α和oxaliplatin明显上调IFNGR2转录水平,并明确了TNF-α通过NF-κB信号通路调控IFNγR2。通过构建荧光素酶报告系统,发现并验证了Ifngr2启动子中NF-κB结合位点。我们还对oxaliplatin对IFNγR2和抗肿瘤免疫的调控作用进行了初步探索。以上研究共同为阐明Ifngr2的转录调控机制打下坚实的基础。
Abstract
gan rao su γ(interferon-γ,IFN-γ)zai xian tian mian yi 、yan zheng fan ying 、kuo ying xing mian yi 、zhong liu mian yi deng sheng ming guo cheng zhong fa hui zhi guan chong yao de zuo yong 。IFN-γjie ge mo biao mian shou ti IFN-γRbing ji huo ji xia you de JAK1、JAK2,jin yi bu lin suan hua zhuai lu yin zi STAT1,STAT1jin ru xi bao he nei jie ge ba ji yin .de te zheng xing GAS(Gamma interferon activation site)yuan jian ,qi dong gan rao su ci ji ji yin (interferon stimulated gene,ISG)de biao da 。ISGzhong bao han you qu hua yin zi 、kang yuan cheng di fen zi 、tun shi shou ti he ge chong ju .you kang .jun kang bing du xiao ying de xi bao yin zi ,can yu xi bao nei bing yuan ti de fang yu ,yan zheng yu mian yi fan ying diao jie ,zhong liu mian yi deng mian yi guo cheng 。IFN-γRyou liang chong ya ji zu cheng :IFNγR1he IFNγR2。IFNγR1biao da xiang dui guo sheng ,IFN-γxin hao tong lu fan ying xing shui ping you IFNγR2de biao da liang jue ding 。yin ci chan ming IFNγR2de diao kong ji zhi you zhu yu shen ru li jie yu IFN-γxin hao tong lu you guan de mian yi fan ying he ji bing fa sheng guo cheng ,ju you qian zai de lin chuang yi yi 。mu qian guan yu IFNγR2de yan jiu zhu yao she ji ji ding wei ji gong neng ,ji zhuai lu diao kong ji zhi yan jiu shao jian bao dao 。ben yan jiu ni tong guo gou jian IFNγR2ying guang bao gao ji tong ,wei fa xian ke diao kong IFNγR2zhuai lu shui ping de ji yin 、xiao fen zi yao wu ,jin yi bu tan suo ji diao kong fang shi he ji ji fen zi ji zhi deng di gong you li gong ju 。wo men jiang xiao shu Ifngr2ji yin de qi dong zi xu lie fang zhi yu lu se ying guang dan bai kai fang yue dou kuang jia shang you ,bing jiang ji gong tong cha ru man bing du zai ti zhong ,jin yi bu tong guo man bing du gan ran ji tong gou jian wen ding biao da Ifngr2qi dong zi diao kong EGFPbiao da de Myc-Capxi bao ji ,yi ci xi bao ji yong zuo yan jiu IFNγR2diao kong de ying guang bao gao ti ji 。wo men fa xian zhong liu huai si yin zi (tumor necrosis factor alpha,TNF-α)he ao sha li bo :(oxaliplatin):ke yi ming xian :shang diao Ifngr2ji yin de zhuai lu shui ping 。TNF-αshi ji ti ji tong xing yan zheng ji xing ji de chong yao xi bao yin zi ,zhu yao you ji huo de ju shi xi bao fen bi ,diao kong ge chong mian yi guo cheng bing ke you dao zhong liu xi bao diao wang 。TNF-αjie ge xi bao biao mian de TNFR,ji huo xia you NF-κBxin hao tong lu he MAPKxin hao tong lu 。wo men fa xian guo biao da NF-κBde ya ji p65ke shang diao Ifngr2。fen bie yong IKK-βyi zhi ji IKK2 inhibitor IV、ML120Bhe dan bai mei ti yi zhi ji MG132yi zhi NF-κBxin hao tong lu shi hou de ji huo ,TNF-αjun bu neng shang diao Ifngr2。zhe biao ming TNF-αtong guo NF-κBxin hao tong lu diao kong Ifngr2,zhe chong diao kong fa sheng zai zhuai lu shui ping 。MAPKxin hao tong lu yi zhi ji U1026、SP100625bu neng zu duan TNF-αcu jin Ifngr2shang diao ,di shi MAPKxin hao tong lu ke neng bu can yu TNF-αshang diao Ifngr2de guo cheng 。wei jin yi bu ming que Ifngr2qi dong zi zhong guan jian de zhuai lu yuan jian ,wo men gou jian xiao shu Ifngr2ji yin qi dong zi ying guang su mei bao gao ji tong ,tong guo ji lie jie duan fen xi tan suo Ifngr2ji yin qi dong zi zhong zhuai lu diao kong de guan jian ou yu 。sheng wu xin xi xue gong ju fen xi fa xian Ifngr2ji yin qi dong zi -111--102pian duan GGGAAAGTCCwei NF-κBjie ge wei dian ,bing tong guo tu bian fen xi dui ji jin hang le que ren 。Oxaliplatinzuo wei yi chong kang zhong liu yao wu ,an fan ying yong yu duo chong zhong liu de hua liao fang an zhong 。zai xi bao shui ping wo men fa xian oxaliplatin.shang diao IFNγR2。wo men jian li le Myc-Cappi xia yi zhi liu dong wu mo xing xing zai dong wu shui ping chu bu tan suo le oxaliplatindui IFNγR2de diao kong zuo yong he kang zhong liu mian yi xiao ying 。ben yan jiu zhong wo men gou jian le IFNγR2ying guang bao gao ti ji ,fa xian le TNF-αhe oxaliplatinming xian shang diao IFNGR2zhuai lu shui ping ,bing ming que le TNF-αtong guo NF-κBxin hao tong lu diao kong IFNγR2。tong guo gou jian ying guang su mei bao gao ji tong ,fa xian bing yan zheng le Ifngr2qi dong zi zhong NF-κBjie ge wei dian 。wo men hai dui oxaliplatindui IFNγR2he kang zhong liu mian yi de diao kong zuo yong jin hang le chu bu tan suo 。yi shang yan jiu gong tong wei chan ming Ifngr2de zhuai lu diao kong ji zhi da xia jian shi de ji chu 。
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论文作者分别是来自军事科学院的王棋，发表于刊物军事科学院2019-09-02论文，是一篇关于干扰素信号通路论文,转录调控论文,军事科学院2019-09-02论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自军事科学院2019-09-02论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：干扰素信号通路论文; 转录调控论文; 军事科学院2019-09-02论文;