Gastro-duodenal fluid induced nuclear factor-κappaB activation and early pre-malignant alterations in murine hypopharyngeal mucosa

// Dimitra P. Vageli 1 , Manju L. Prasad 2 and Clarence T. Sasaki 1 1 Department of Surgery,Yale Larynx Laboratory Section of Otolaryngology, Yale School of Medicine, New Haven, CT, USA 2 Pathology and of Surgery (Otolaryngology), Yale School of Medicine, New Haven, CT, USA Correspondence to: Dimitra P. Vageli, email: // Keywords : hypopharyngeal cancer, NF-κB, gastroduodenal reflux, bile acids, in vivo Received : October 19, 2015 Accepted : December 24, 2015 Published : January 06, 2016 Abstract We recently described the role of gastro-duodenal fluids (GDFs) in generating changes consistent with hypopharyngeal neoplasia through activation of NF- κ B pathway, using an in vitro model of human hypopharyngeal normal keratinocytes. Here, we further provide evidence that gastro-duodenal reflux is a risk factor for early pre-malignant alterations in hypopharyngeal mucosa (HM) related to an activated NF- κ B oncogenic pathway, using both an in vitro and a novel in vivo model of C57Bl/6J mice. Histological, immunohistochemical and automated quantitative analysis documents significant NF- κ B activation and early pre-malignant alterations in HM topically exposed to GDFs, compared to acid alone and other controls. Early pre-malignant histologic lesions exhibited increased Ki67, CK14 and ΔNp63, cell proliferation markers, changes of cell adhesion molecules, E-Cadherin and β-catenin, and STAT3 activation. The in vivo effect of NF- κ B activation is positively correlated with p-STAT3, Ki67, CK14 or β-catenin expression, while GDFs induce significant transcriptional activation of RELA(p65), bcl-2, TNF-α, STAT3, EGFR and wnt5A, in vivo . Our in vivo model demonstrates selectively activated NF- κ B in response to topically administrated GDFs, leading to early pre-malignant events in HM.