华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

脾络氨酸激酶-核因子κB调控口腔癌相关巨噬细胞中癌痛相关环氧化酶2的机制研究

林洁1 王淼1 吉阳1 刘乐1 高攀 李春洁2   

  1. 1.口腔疾病研究国家重点实验室 华西口腔医院口腔麻醉科(四川大学);2.头颈肿瘤外科,成都 610041
  • 收稿日期:2016-03-16 修回日期:2016-07-12 出版日期:2016-10-01 发布日期:2016-10-01
  • 通讯作者: 李春洁,副教授,博士,E-mail:lichunjie@scu.edu.cn
  • 作者简介:林洁,主治医师,硕士,E-mail:514541402@qq.com
  • 基金资助:

    2014年四川大学青年教师科研启动基金(2014SCU11032);2015年四川大学华西口腔医院青年科学研究基金(2015-06)

In vitro investigation on the mechanism of cyclooxygenase-2 upregulation induced by spleen tyrosine kinase-nuclear factor κB signaling in cancer pain caused by oral cancer-associated macrophag

Lin Jie1, Wang Miao1, Ji Yang1, Liu Le1, Gao Pan2, Li Chunjie2.   

  1. 1. State Key Laboratory of Oral Diseases, Dept. of Dental Anesthesia, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China; 2. State Key Laboratory of Oral Diseases, Dept. of Head and Neck Oncology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2016-03-16 Revised:2016-07-12 Online:2016-10-01 Published:2016-10-01
  • Contact: Li Chunjie, E-mail: lichunjie@scu.edu.cn.
  • Supported by:

    Scientific Research Foundation for Young Teachers of Sichuan University (2014SCU11032); Scientific Research Foundation for Young Investigators of West China Hospital of Stomatology, Sichuan University (2015-06).

摘要:

目的 通过体外原代巨噬细胞诱导以及分子生物学的方法,探究口腔癌相关巨噬细胞环氧化酶2(COX-2)上调的机制。方法  构建小鼠原代巨噬细胞,使用Cal27条件培养基(CM)刺激诱导形成口腔癌相关巨噬细胞,使用免疫荧光检测原代巨噬细胞的纯度。使用小分子抑制剂分别抑制脾络氨酸激酶(Syk)及核因子κB(NFκB)通路。使用实时定量聚合酶链反应(PCR)、Western blot检测COX-2及信号通路相关蛋白的变化。结果  所诱导的原代巨噬细胞均特异性表达CD68蛋白。Cal27 CM刺激能够显著提高COX-2的表达(P<0.001);抑制Syk的磷酸化即能够进一步抑制NFκB-P65的磷酸化,从而导致COX-2的表达显著降低(P<0.01);而抑制NFκB-P65的磷酸化不能抑制Syk的磷酸化但可以显著降低COX-2的表达(P<0.01)。结论  Syk-NFκB信号通路导致COX-2在口腔癌相关的巨噬细胞中高表达,靶定该信号通路可能是控制口腔癌相关癌痛的新方向。

关键词: 癌痛, 口腔癌, 环氧化酶2, 巨噬细胞

Abstract:

Objective  This study explores the mechanism of cyclooxygenase-2 (COX-2) upregulation in oral cancers asso- ciated with macrophage by using molecular biology techniques and primary culture of murine macrophage. Methods  Murine macrophage was induced by macrophage colony-stimulating factor (M-CSF) and Cal27 conditional medium (CM). Purity of the macrophage was detected through CD68 immunofluorescence staining. Inhibitors of spleen tyrosine kinase (Syk) and nuclear factor κB (NFκB) were used to inhibit these pathways. In addition, real-time polymerase chain reaction and Western blot analysis were used to detect alterations in COX-2 and pathway-related proteins. Results  All of the induced cells specifically expressed CD68. Cal27 CM could significantly induce COX-2 expression (P<0.001). Moreover, inhibition of Syk pathway attenuated NFκB-P65 phosphorylation and reduced COX-2 expression (P<0.01), and inhibition of NFκB pathway exerted no effects on Syk phosphorylation but significantly inhibited COX-2 upregulation (P<0.01). Conclusion  Syk-NFκB is responsible for COX-2 overexpression in oral cancer associated with macrophages. Targeting this pathway is possibly a new approach to control oral cancer-related pain.

Key words: cancer pain, mouth neoplasm, cyclooxy- genase-2, macrophage