Cyclooxygenase-2 signaling in vocal fold fibroblasts.
OBJECTIVE
Inflammation and its role in a coordinated fibroplastic response, which disrupts the structure of the vocal folds following injury, is critical. Cyclooxygenase-2 (COX-2) is an important enzyme involved in both inflammation and fibrosis; in addition, it is a prime target for therapeutic intervention. We sought to study this pathway in vocal fold fibroblasts to provide a foundation for future interventional studies.


METHODS
In vitro.


METHODS
Human vocal fold fibroblasts were incubated with IL-1 beta to determine the effects on COX-2 signaling, along with upstream regulatory mechanisms and downstream mediators of wound healing. In vitro methods to assess mRNA expression, as well as intracellular and secreted protein (sodium dodecyl sulfate polyacrylamide gel electrophoresis and enzyme-linked immunosorbent assay) were employed.


RESULTS
IL-1 beta regulation of COX-2 mRNA and protein levels was dose and time dependent and IL-1 beta altered PGE(2) metabolism, via regulation of both synthetic and degradative enzymes. IL-1 beta increased nuclear factor (NF)-kappaB activation and nuclear translocation. Inhibition of the p50 and p65 subunits of NF-kappaB decreased IL-1 beta-induced COX-2 transcription. IL-1 beta also altered mRNA expression of four cell-surface prostaglandin receptors.


CONCLUSIONS
Inflammation and fibrosis are important in the vocal fold pathophysiologic response to injury. Our data suggest that COX-2 and PGE(2) are inducible in human vocal fold fibroblasts, and this response appears to be NF-kappaB-dependent. We purport this fundamental investigation will lead to increased insight regarding injury and repair in the vocal folds, with the ultimate goal of developing novel clinical care paradigms.
Branski
Ryan C
RC
Department of Head and Neck Surgery, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA. branskir@mskcc.org
Zhou
Hang
H
Sandulache
Vlad C
VC
Chen
Jie
J
Felsen
Diane
D
Kraus
Dennis H
DH
D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000235:genetics*; D015232:Dinoprostone / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D053583:Interleukin-1beta / Q000494:pharmacology*; D016328:NF-kappa B / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics; D011982:Receptors, Prostaglandin / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects* / Q000235:genetics*; D014158:Transcription, Genetic; D014827:Vocal Cords / Q000187:drug effects*
D053583:Interleukin-1beta; D016328:NF-kappa B; D012333:RNA, Messenger; D011982:Receptors, Prostaglandin; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D015232:Dinoprostone