[1]孙锦霞,黄钟,王瑞,等.巨噬细胞中FBW7沉默对其杀菌能力的抑制[J].深圳大学学报理工版,2017,34(No.4(331-440)):358-363.[doi:10.3724/SP.J.1249.2017.04358]
 Sun Jinxia,Huang Zhong,Wang Rui,et al.The anti-bactericidal effects of FBW7 silence in macrophage[J].Journal of Shenzhen University Science and Engineering,2017,34(No.4(331-440)):358-363.[doi:10.3724/SP.J.1249.2017.04358]
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巨噬细胞中FBW7沉默对其杀菌能力的抑制()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第34卷
期数:
2017年No.4(331-440)
页码:
358-363
栏目:
生物工程
出版日期:
2017-07-10

文章信息/Info

Title:
The anti-bactericidal effects of FBW7 silence in macrophage
文章编号:
201704004
作者:
孙锦霞1黄钟1王瑞2翁林军3李亚旭3
1)深圳大学医学部,广东深圳518060
2)聊城大学药学院,山东聊城252059
3)同济大学生命科学与技术学院,上海200092
Author(s):
Sun Jinxia1 Huang Zhong1 Wang Rui2 Weng Linjun3 and Li Yaxu3
1) Health Science Center, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China
2) College of Pharmacy, Liaocheng University, Liaocheng 252059, Shandong Province, P.R.China
3) School of Life Science and Technology, Tongji University, Shanghai 200092, P.R.China
关键词:
小干扰核糖核酸RAW264.7细胞巨噬细胞杀菌活性一氧化氮合成酶NF-κB信号通路实时荧光定量聚合酶链式反应
Keywords:
small interfering RNA (siRNA) RAW264.7 macrophage bactericidal activity nitric oxide synthase (iNOS) NF-κB signaling pathway real time quantitative polymerase chain reaction (RT-qPCR)
分类号:
R 392
DOI:
10.3724/SP.J.1249.2017.04358
文献标志码:
A
摘要:
采用小干扰核糖核酸(siRNA)介导Hela细胞、原代巨噬细胞和RAW264.7细胞中FBW7基因沉默,研究其对NF-κB信号通路、细胞因子表达、巨噬细胞吞噬和杀伤细菌能力的影响.通过流式细胞术、抗生素保护试验、实时荧光定量聚合酶链式反应和报告基因的研究方法,发现FBW7沉默不具有调控RAW264.7细胞吞噬细菌的能力,但能显著抑制原代巨噬细胞和RAW264.7细胞的杀菌能力及一氧化氮合成酶(nitric oxide synthase, iNOS)表达,抑制Hela中NF-κB报告基因活性,降低Hela细胞中TNFα、 IL-1β和IL-6, 以及原代巨噬细胞、RAW264.7细胞中MCP-1和IL-1β表达.结果表明,FBW7沉默可通过影响NF-κB活性抑制巨噬细胞对细菌的杀伤能力.
Abstract:
SiRNA-mediated FBW7 silence in Hela, primary macrophage and RAW264.7 were performed to assay the role of FBW7 on NF-κB activity, cytokines expression and phagocytosis and bactericidal activity of macrophage. Flow cytometry (FCM), antibiotic protection assay (APA), real time quantitative polymerase chain reaction (RT-qPCR) and report gene analysis suggest that FBW7 silence does not affect phagocytosis of RAW264.7, but significantly inhibits bactericidal activity and nitric oxide synthase (iNOS) expression in primary macrophage and RAW264.7, suppresses NF-κB report gene activity in Hela, decreases expression of TNFα、 IL-1β、 IL-6 in Hela and MCP-1、 IL-1β in primary macrophage and RAW264.7. Thus, FBW7 silence might inhibit bactericidal activity of macrophage by regulating NF-κB activity.

参考文献/References:

[1] Scallan E, Griffin P M, Angulo F J, et al. Foodborne illness acquired in the United States—unspecified agents[J]. Emerging Infectious Diseases, 2011, 17(1): 16-22.
[2] Murray P J, Wynn T A. Protective and pathogenic functions of macrophage subsets[J]. Nature Reviews Immunology, 2011, 11(11): 723-737.
[3] Wynn T A, Chawla A, Pollard J W. Macrophage biology in development, homeostasis and disease[J]. Nature, 2013, 496(7446): 445-455.
[4] Cao Jun, Ge Minghua, Ling Zhiqiang. Fbxw7 tumor suppressor: a vital regulator contributes to human tumorigenesis[J]. Medicine (Baltimore), 2016, 95(7): e2496.
[5] Rahman M M, McFadden G. Modulation of NF-κB signalling by microbial pathogens[J]. Nature Reviews Microbiology, 2011, 9(4): 291-306.
[6] MacMicking J, Xie Qiaowen, Nathan C. Nitric oxide and macrophage function[J]. Annual Review of Immunology, 1997, 15(15): 323-350.
[7] Nairz M, Schleicher U, Schroll A, et al. Nitric oxide-mediated regulation of ferroportin-1 controls macrophage iron homeostasis and immune function in Salmonella infection[J]. The Journal of Experimental Medicine, 2013, 210(5): 855-873.
[8] Tu Kangsheng, Zheng Xin, Zhou Zhenyu, et al. Recombinant human adenovirus-p53 injection induced apoptosis in hepatocellular carcinoma cell lines mediated by p53-Fbxw7 pathway, which controls c-Myc and cyclin E[J]. PLOS One, 2013, 8(7): e68574.
[9] Yokobori T, Mimori K, Iwatsuki M, et al. Copy number loss of FBXW7 is related to gene expression and poor prognosis in esophageal squamous cell carcinoma[J]. International Journal of Oncology, 2012, 41(1): 253-259.
[10] Gao Jiankun, Azmi A S, Aboukamee A, et al. Nuclear retention of Fbw7 by specific inhibitors of nuclear export leads to Notch1 degradation in pancreatic cancer[J]. Oncotarget, 2014, 5(11): 3444-3454.
[11] Aydin I T, Melamed R D, Adams S J, et al. FBXW7 mutations in melanoma and a new therapeutic paradigm[J]. Journal of the National Cancer Institute, 2014, 106(6): dju107.
[12] Luan Yi, Wang Ping. FBW7-mediated ubiquitination and degradation of KLF5[J]. World Journal of Biological Chemistry, 2014, 5(2): 216-223.
[13] Wang Rui, Wang Yan, Liu Ning, et al. FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation[J]. Cell Research, 2013, 23(6): 803-819.
[14] Arabi A, Ullah K, Branca R M M, et al. Proteomic screen reveals Fbw7 as a modulator of the NF-κB pathway[J]. Nature Communications, 2012, 3: 976.
[15] Fukushima H, Matsumoto A, Inuzuka H, et al. SCFFbw7 modulates the NFκB signaling pathway by targeting NFkB2 for ubiquitination and destruction[J]. Cell Reports, 2012, 1(5): 434-443.
[16] Huang Haishan, Ma Li, Li Jingxia, et al. NF-κB1 inhibits c-Myc protein degradation through suppression of FBW7 expression[J]. Oncotarget, 2014, 5(2): 493-505.
[17] Balamurugan K, Sharan S, Klarmann K D, et al. FBXW7α attenuates inflammatory signalling by downregulating C/EBPδ and its target gene Tlr4[J]. Nature Communications, 2013, 4: 1662.

备注/Memo

备注/Memo:
Received:2016-11-05;Revised:2017-02-28;Accepted:2017-05-07
Foundation:National Natural Science Foundation of China (31401217); Postdoctral Science Foundation of China (2014M0560672)
Corresponding author:Professor Huang Zhong. E-mail: zhuang809@126.com
Citation:Sun Jinxia, Huang Zhong, Wang Rui, et al. The anti-bactericidal effects of FBW7 silence in macrophage[J]. Journal of Shenzhen University Science and Engineering, 2017, 34(4): 358-363.(in Chinese)
基金项目:国家自然科学基金资助项目(31401217);中国博士后科学基金资助项目(2014M0560672)
作者简介:孙锦霞(1986—),女,深圳大学博士后研究人员. 研究方向:免疫学. E-mail:jinxia8608@126.com
引文:孙锦霞,黄钟,王瑞,等. 巨噬细胞中FBW7沉默对其杀菌能力的抑制[J]. 深圳大学学报理工版,2017,34(4):358-363.
更新日期/Last Update: 2017-06-26