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

Cancer. 2019 Oct 15;125(20):3554-3565. doi: 10.1002/cncr.32369. Epub 2019 Jul 16.

Abstract

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.

Keywords: NF-κB; bile reflux; hypopharyngeal cancer; miR-21; miR-375.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Animals
  • Bile / metabolism
  • Bile Acids and Salts / metabolism
  • Bile Acids and Salts / toxicity
  • Bile Reflux / complications
  • Bile Reflux / genetics*
  • Bile Reflux / metabolism
  • Bile Reflux / pathology
  • Carcinoma, Squamous Cell / complications
  • Carcinoma, Squamous Cell / genetics*
  • Carcinoma, Squamous Cell / metabolism
  • Carcinoma, Squamous Cell / pathology
  • Female
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Hypopharyngeal Neoplasms / complications
  • Hypopharyngeal Neoplasms / genetics*
  • Hypopharyngeal Neoplasms / metabolism
  • Hypopharyngeal Neoplasms / pathology
  • Male
  • Mice
  • MicroRNAs / genetics
  • Middle Aged
  • Mucous Membrane / drug effects
  • Mucous Membrane / pathology
  • NF-kappa B / genetics
  • Neoplasm Proteins / genetics*
  • RNA, Messenger / genetics

Substances

  • Bile Acids and Salts
  • MIRN21 microRNA, mouse
  • MicroRNAs
  • Mirn375 microRNA, mouse
  • NF-kappa B
  • Neoplasm Proteins
  • RNA, Messenger