Biliary reflux as a causal factor in hypopharyngeal carcinoma: New clinical evidence and implications

• Published in: Cancer 2019-10, Vol.125 (20), p.3554-3565
• Main Authors: Sasaki, Clarence T., Doukas, Sotirios G., Costa, Jose, Vageli, Dimitra P.
• Format: Article
• Language: English
• Subjects: Aged; Animals; Ants; Bile; Bile - metabolism; Bile Acids and Salts - metabolism; Bile Acids and Salts - toxicity; bile reflux; Bile Reflux - complications; Bile Reflux - genetics; Bile Reflux - metabolism; Bile Reflux - pathology; Carcinogenesis; Carcinogens; Carcinoma, Squamous Cell - complications; Carcinoma, Squamous Cell - genetics; Carcinoma, Squamous Cell - metabolism; Carcinoma, Squamous Cell - pathology; Disease control; Epidermal growth factor receptors; Esophagus; Female; Gene Expression Regulation, Neoplastic; Health risks; Humans; hypopharyngeal cancer; Hypopharyngeal Neoplasms - complications; Hypopharyngeal Neoplasms - genetics; Hypopharyngeal Neoplasms - metabolism; Hypopharyngeal Neoplasms - pathology; Interleukin 1; Male; Mice; MicroRNAs; MicroRNAs - genetics; Middle Aged; miRNA; miR‐21; miR‐375; mRNA; Mucosa; Mucous Membrane - drug effects; Mucous Membrane - pathology; Neoplasm Proteins - genetics; NF-kappa B - genetics; NF‐κB; Notch1 protein; Oncology; p53 Protein; Phenotypes; Polymerase chain reaction; RelA protein; Ribonucleic acid; Risk analysis; Risk factors; RNA; RNA, Messenger - genetics; Squamous cell carcinoma; Stat3 protein; Throat cancer; Tumor necrosis factor; Tumor suppressor genes; Tumors; Wnt protein
• ISSN: 0008-543X; 1097-0142
• DOI: 10.1002/cncr.32369

Background Recent preclinical explorations strongly support the tumorigenic potential of bile on laryngopharyngeal mucosa. Herein, the authors describe, in bile‐related human hypopharyngeal squamous cell carcinoma (HSCC), NF‐κB–related messenger RNA (mRNA) and microRNA (miRNA) oncogenic phenotypes similar to those previously identified in acidic bile–exposed premalignant murine hypopharyngeal mucosa. Methods In this pilot study, the authors included human HSCC specimens paired with their adjacent normal tissue (ANT) derived from 3 representative patients with documented biliary laryngopharyngeal reflux (bile[+]) compared with 5 control patients without signs of bile reflux disease (bile[‐]). Immunohistochemical, quantitative polymerase chain reaction, and miRNA analyses were used to detect the levels of activated NF‐κB and expression levels of STAT3, EGFR, BCL2, WNT5A, IL‐6, IL‐1B, ΔNp63, cREL, TNF‐α, TP53, NOTCH1, NOTCH2, NOTCH3, miR‐21, miR‐155, miR‐192, miR‐34a, miR‐375, miR‐451a, miR‐489, miR‐504, and miR‐99a. Results Bile(+) HSCC demonstrated an intense NF‐κB activation accompanied by significant overexpression of RELA(p65), EGFR, STAT3, BCL‐2, cREL, ΔNp63, WNT5A, IL‐6, and IL1B; upregulation of oncomir miR‐21; and downregulation of tumor suppressor miR‐375 compared with their respective ANTs. Bile(+) HSCC demonstrated significantly higher mRNA levels of all the analyzed genes, particularly RELA(p65), IL‐6, EGFR, and TNF‐α compared with bile(‐) tumors. The miR‐21/miR‐375 ratio, which previously has been linked to tumor aggressiveness, was found to be >260‐fold and >30‐fold higher, respectively, in bile(+) HSCCs compared with their ANTs and bile(‐) tumors. Conclusions Although limitations apply to this pilot study due to the small number of patients with HSCC, the novel findings suggest that a history of bile as a component of esophageal reflux disease may represent an independent risk factor for hypopharyngeal carcinogenesis. The current pilot study provides evidence suggesting that bile has a pathogenetic role in the genesis of hypopharyngeal cancer. A subset of hypopharyngeal carcinomas in patients with bile reflux demonstrate a characteristic molecular profile that distinguishes them from carcinomas arising in patients with no evidence of bile reflux, and can be used as a tool for patient stratification in possible targeted therapies.