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Pro-inflammatory Cytokines Up-regulate MUC1 Gene Expression in Oral Epithelial Cells

¹ Departments of Periodontology and Oral Biology,
² Medicine, and
³ Biochemistry, GI Section, X-510, 650 Albany Street, Boston University Medical Center, Boston, MA 02118, USA;

View larger version (32K): [in this window] [in a new window]   Figure 1. Expression of MUC1 in KB oral epithelial cells. (Lane 1) Northern blot of RNA isolated from KB cells hybridized with a 32P-labeled MUC1 tandem-repeat probe. The positions of the 6.4- and 4.7-kb MUC1 transcripts are indicated with arrows. (Lanes 2, 3) Western blot of proteins in KB cell lysates probed with MUC1-specific antibodies. Blots were probed with monoclonal antibody VU-4H5 (1:500 dilution) to detect the extracellular subunit of MUC1 (lane 2) or with monoclonal antibody CT2 (1:300 dilution) to detect the cytoplasmic subunit of MUC1 (lane 3).

View larger version (15K): [in this window] [in a new window]   Figure 2. Pro-inflammatory cytokines and oral bacteria up-regulate MUC1 mRNA expression. RNA was isolated from control cells and cells treated with cytokines, LPS, or P. gingivalis for 4 hrs and reverse-transcribed; the resulting cDNAs were amplified by real-time PCR with MUC1- and β-actin-specific primers. Transcript levels were calculated by reference to the standard curves, and MUC1 mRNA levels were normalized to those of β-actin. Each bar represents cDNA from 3 independent experiments analyzed in triplicate. The fold increase in transcript levels over untreated controls is expressed as mean ± SEM, and statistical significance (P < 0.05) is indicated with an asterisk.

View larger version (34K): [in this window] [in a new window]   Figure 3. Pro-inflammatory cytokines up-regulate MUC1 protein expression. Lysates from control cells and cells treated with combinations of cytokines for 8 hrs were subjected to electrophoresis on Tris-tricine gels and blotted; the blots were probed with the MUC1 antibody CT2 as described in the legend to Fig. 1.

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