Tumor and Stem Cell Biology D Np 63 Versatilely Regulates a Broad NFk B Gene Program and Promotes Squamous Epithelial Proliferation , Migration , and Inflammation

Head and neck squamous cell carcinoma (HNSCC) and many other epithelial malignancies exhibit increased proliferation, invasion, and inflammation, concomitant with aberrant nuclear activation of TP53 and NF-kB family members DNp63, cRel, and RelA. However, the mechanisms of cross-talk by which these transcription factors coordinate gene expression and the malignant phenotype remain elusive. In this study, we showed that DNp63 regulates a cohort of genes involved in cell growth, survival, adhesion, and inflammation, which substantially overlaps with the NF-kB transcriptome. DNp63 with cRel and/or RelA are recruited to form novel binding complexes on p63 or NF-kB/Rel sites of multitarget gene promoters. Overexpressed DNp63or TNF-a–induced NF-kB and inflammatory cytokine interleukin-8 (IL-8) reporter activation depended on RelA/cRel regulatory binding sites. Depletion of RelA or DNp63 by small interfering RNA (siRNA) significantly inhibited NF-kB–specific, or TNF-a–induced IL-8 reporter activation. DNp63 siRNA significantly inhibited proliferation, survival, and migration by HNSCC cells in vitro. Consistent with these data, an increase in nuclear DNp63, accompanied by increased proliferation (Ki-67) and adhesion (b4 integrin) markers, and induced inflammatory cell infiltration was observed throughout HNSCC specimens, when compared with the basilar pattern of protein expression and minimal inflammation seen in nonmalignant mucosa. Furthermore, overexpression of DNp63a in squamous epithelial cells in transgenic mice leads to increased suprabasilar cRel, Ki-67, and cytokine expression, together with epidermal hyperplasia and diffuse inflammation, similar to HNSCC. Our study reveals DNp63 as a master transcription factor that, in coordination with NF-kB/Rels, orchestrates a broad gene program promoting epidermal hyperplasia, inflammation, and the malignant phenotype of HNSCC. Cancer Res; 71(10); 3688–700. 2011 AACR.

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