Inhibition of LPS-induced cyclooxygenase 2 and nitric oxide production by transduced PEP-1-PTEN fusion protein in Raw 264.7 macrophage cells.
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.
Lee
Sun Hwa
SH
Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, College of Medicine, Hallym University, Chunchon 200-702, Korea.
Lee
Yeom Pyo
YP
Kim
So Young
SY
Jeong
Min Seop
MS
Lee
Min Jung
MJ
Kang
Hye Won
HW
Jeong
Hoon Jae
HJ
Kim
Dae Won
DW
Sohn
Eun Joung
EJ
Jang
Sang Ho
SH
Kim
Yeon Hyang
YH
Kwon
Hyung Joo
HJ
Cho
Sung Woo
SW
Park
Jinseu
J
Eum
Won Sik
WS
Choi
Soo Young
SY
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
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