1. Tec kinase mediating IL-8 transcription in monocytes stimulated with LPS
- Author
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Guang-Qing Wang, Yi-Tao Jia, Zhaofan Xia, and Xiao-Yan Yang
- Subjects
Lipopolysaccharides ,Transcription, Genetic ,education ,Immunology ,Mitogen-activated protein kinase kinase ,Biology ,Monocytes ,Cell Line ,Mice ,Mediator ,Transcription (biology) ,medicine ,Immunology and Allergy ,Animals ,Point Mutation ,Luciferase ,Interleukin 8 ,Promoter Regions, Genetic ,Monocyte ,Macrophages ,Interleukin-8 ,hemic and immune systems ,Transfection ,Protein-Tyrosine Kinases ,Molecular biology ,medicine.anatomical_structure ,Cell culture ,tissues - Abstract
The purpose of present study was to explore the possibility of Tec kinase as a mediator for IL-8 transcription in monocytes stimulated with LPS. Plasmids of mouse Tec kinase IV or Tec kinase IV with inactivating point mutations generated with QuikChange site-directed mutagenesis were co-transfected with IL-8 promoter driven luciferase construct into RAW264.7 cells, then luciferase activity was measured with a luminometer. The results shown Tec kinase could significantly enhance IL-8 transcription. Furthermore, point inactivating mutation in SH2, PH or PTK domain almost completely abolish the effects of Tec kinase on the transcription of IL-8. In the transfection experiment, PD98059, a MEK1 inhibitor, decreased the transcription of IL-8 in a dose dependent pattern. When siRNA for Tec kinase was transfected into THP-1 cells, it could efficiently block the production of IL-8 from THP-1 cells (p < 0.01) stimulated with LPS. In conclusion, Tec kinase may mediate the transcription of IL-8 in monocyte stimulated with LPS.
- Published
- 2009