Tumor necrosis factor increases MUC1 mRNA in cultured human nasal epithelial cells.
OBJECTIVE
Mucins are high molecular weight glycoproteins which are normally expressed on the surface of a variety of epithelia. It is possible that shedding of such molecules from the epithelium could play a role in preventing bacterial colonization at the mucosal surface. Immunohistochemical and reverse transcriptase polymerase chain reaction(RT-PCR) analyses of human inferior turbinates have shown the existence of MUC1 mucin in nasal mucosa. However, the regulatory mechanisms of MUC1 mucin are poorly understood. In order to clarify the modulation of mucin gene expression, we developed a real-time semi-quantitative RT-PCR based on TaqMan fluorescence methodology to quantify MUC1 mRNA in primary cultured human nasal epithelial cells (HNECs).


METHODS
HNECs were stimulated with recombinant human tumor necrosis factor (TNF)-alpha (20 pg/ml to 20 ng/ml) for specified time periods (0, 12, 24 and 48 h) and MUC1 mRNA was determined by means of semi-quantitative RT-PCR.


RESULTS
Significant increases in MUC1 gene expression in HNECs were initially detected at 12 h, peaking at 24 h after stimulation. TNF-mediated MUCI mRNA expression at 24 h was significantly inhibited by co-incubation with human recombinant soluble TNF receptor.


CONCLUSIONS
TNF-mediated MUC1 gene expression may contribute to the pathogenesis of human inflammatory upper airway disorders. Also, our mucin mRNA real-time PCR provides a quantitative method for investigating the regulation of mucin gene expression in both healthy and diseased samples.
Shirasaki
Hideaki
H
Department of Otolaryngology, Sapporo Medical University, Sapporo, Japan. puchibe@spa.att.ne.jp
Kanaizumi
Etsuko
E
Watanabe
Kazumasa
K
Konno
Nobuhiro
N
Sato
Jun
J
Narita
Shin-Ichirou
S
Himi
Tetsuo
T
D000328:Adult; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008875:Middle Aged; D018396:Mucin-1 / Q000096:biosynthesis* / Q000235:genetics; D009297:Nasal Mucosa / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology* / Q000502:physiology
D018396:Mucin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha