Exp Mol Med.
2008 Dec;40(6):629-638. 10.3858/emm.2008.40.6.629.
Inhibition of LPS-induced cyclooxygenase 2 and nitric oxide production by transduced PEP-1-PTEN fusion protein in Raw 264.7 macrophage cells
- Affiliations
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- 1Department of Biomedical Science and Research Institute for Bioscience and Biotechnology,College of Medicine, Hallym University, Chunchon, Korea. sychoi@hallym.ac.kr, wseum@hallym.ac.kr
- 2Department of Microbiolgy, College of Medicine, Hallym University, Chunchon, Korea.
- 3Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine, Seoul, Korea.
- KMID: 1072943
- DOI: http://doi.org/10.3858/emm.2008.40.6.629
Abstract
- Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) is a tumor suppressor. Although it is well known to have various physiological roles in cancer, its inhibitory effect on inflammation remains poorly understood. In the present study, a human PTEN gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-PTEN fusion protein. The expressed and purified PEP-1-PTEN fusion protein were transduced efficiently into macrophage Raw 264.7 cells in a time- and dose- dependent manner when added exogenously in culture media. Once inside the cells, the transduced PEP-1-PTEN protein was stable for 24 h. Transduced PEP-1-PTEN fusion protein inhibited the LPS-induced cyclooxygenase 2 (COX-2) and iNOS expression levels in a dose-dependent manner. Furthermore, transduced PEP-1-PTEN fusion protein inhibited the activation of NF-kappa B induced by LPS. These results suggest that the PEP-1-PTEN fusion protein can be used in protein therapy for inflammatory disorders.
MeSH Terms
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Animals
Cell Line
Cyclooxygenase 2/*metabolism
Cysteamine/*analogs & derivatives
Enzyme Activation
Humans
Lipopolysaccharides/*pharmacology
Macrophages/*metabolism
Mice
NF-kappa B/metabolism
Nitric Oxide/*biosynthesis
Nitric Oxide Synthase Type II/metabolism
PTEN Phosphohydrolase/*genetics
Peptides/*genetics
Recombinant Fusion Proteins/*biosynthesis/genetics
Signal Transduction
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