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B is an Important Modulator of the Altered Gene Expression Profile and Malignant Phenotype in Squamous Cell Carcinoma
1 Head and Neck Surgery Branch, National Institute on Deafness and Other Communication Disorders, Rockville, Maryland; 2 Radiation Oncology Sciences Program, National Cancer Institute, Bethesda, Maryland; and 3 Lineberger Cancer Center, University of North Carolina, Chapel Hill, North Carolina
We reported previously that transcription factor nuclear factor (NF)-
B is constitutively activated in human and murine squamous cell carcinomas (SCCs). The role of NF-B in the cumulative changes in gene expression with transformation and progression of the murine SCC Pam 212 and after switching off NF-B by a dominant negative inhibitor B mutant (IB
M) was explored by profiling with a 15,000-element cDNA micoarrray. Remarkably, NF-B modulated the expression of >60% of the 308 genes differentially expressed between normal keratinocytes and metastatic SCCs. NF-B directly or indirectly modulated expression of programs of genes functionally linked to proliferation, apoptosis, adhesion, and angiogenesis. Among these, changes in expression of cyclin D1, inhibitor of apoptosis-1, mutant Trp53, and ß-catenin detected with modulation of NF-B by microarray were confirmed by Western and Northern blot. NF-B DNA binding motifs were detected in the promoter of 
63% of genes showing increased expression and 33% of the genes showing decreased expression. The ACTACAG motif implicated in the NF-B-dependent down-regulation of mRNA expression of MyoD and Sox9 was detected in the coding portion of about 15% of genes showing increased or decreased expression. Inactivation of NF-B inhibited malignant phenotypic features including proliferation, cell survival, migration, angiogenesis, and tumorigenesis. These results provide evidence that NF-B is an important modulator of gene expression programs that contribute to the malignant phenotype of SCC.
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