title: Inhibition of LPS-induced cyclooxygenase 2 and nitric oxide production by transduced PEP-1-PTEN fusion protein in Raw 264.7 macrophage cells.
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-kappaB induced by LPS. These results suggest that the PEP-1-PTEN fusion protein can be used in protein therapy for inflammatory disorders.
authors.affiliation: Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University, Chunchon 200-702, Korea.
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Lee
authors.forename: Yeom Pyo
authors.initials: YP
authors.lastname: Lee
authors.forename: So Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Min Seop
authors.initials: MS
authors.lastname: Jeong
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Lee
authors.forename: Hye Won
authors.initials: HW
authors.lastname: Kang
authors.forename: Hoon Jae
authors.initials: HJ
authors.lastname: Jeong
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Eun Joung
authors.initials: EJ
authors.lastname: Sohn
authors.forename: Sang Ho
authors.initials: SH
authors.lastname: Jang
authors.forename: Yeon Hyang
authors.initials: YH
authors.lastname: Kim
authors.forename: Hyung Joo
authors.initials: HJ
authors.lastname: Kwon
authors.forename: Sung Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Eum
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000818:Animals; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D003543:Cysteamine / Q000031:analogs & derivatives*; D004789:Enzyme Activation; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D051059:PTEN Phosphohydrolase / Q000235:genetics*; D010455:Peptides / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D015398:Signal Transduction
chemical_list: D008070:Lipopolysaccharides; D016328:NF-kappa B; C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D009569:Nitric Oxide; D003543:Cysteamine; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human