Characterization of functional transient receptor potential melastatin 8 channels in human pancreatic ductal adenocarcinoma cells

Pancreas. 2014 Jul;43(5):795-800. doi: 10.1097/MPA.0000000000000106.

Abstract

Objective: Recently, the transient receptor potential melastatin 8 (TRPM8) channel has emerged as a putative biomarker for pancreatic ductal adenocarcinoma (PDA). This study aimed to evaluate the expression of TRPM8 and its modulation by specific agonists and antagonists in PDA cells.

Methods: We examined the protein expression of TRPM8 in 3 different PDA cell lines and compared it with a nontumoral epithelial cell line of human pancreatic origin using Western blotting and immunocytochemical analysis. To assess the function of TRPM8 channels, we measured the TRPM8 currents in whole-cell mode of the patch clamp technique. To explore the putative involvement of TRPM8 in cell migration, we investigated the motility of PDA cells using the scratch-wound assay.

Results: Pancreatic ductal adenocarcinoma cells express functional plasma membrane TRPM8 channels, which are responsive after exposure to agonists (menthol and icilin) and antagonists N-(3-aminopropyl)-2-{[(3-methylphenyl) methyl]oxy}-N-(2-thienylmethyl)benzamide hydrochloride salt. The silencing of TRPM8 expression by small interfering RNA augments the migration of PDA cells. Conversely, the activated form of TRPM8 inhibits PDA cell motility.

Conclusions: An unglycosylated TRPM8 protein is expressed and is functional in the membrane of PDA cells. Transient receptor potential melastatin 8 inhibits the migration of PDA cells, suggesting a putative role as a biomarker or target for this channel for PDA therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / genetics
  • Adenocarcinoma / metabolism
  • Adenocarcinoma / pathology
  • Benzamides / pharmacology
  • Blotting, Western
  • Carcinoma, Pancreatic Ductal / genetics
  • Carcinoma, Pancreatic Ductal / metabolism
  • Carcinoma, Pancreatic Ductal / pathology
  • Cell Line
  • Cell Line, Tumor
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Cell Membrane / physiology
  • Cell Movement / drug effects
  • Cell Movement / genetics
  • HEK293 Cells
  • Humans
  • Immunohistochemistry
  • Membrane Potentials / drug effects
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Menthol / pharmacology
  • Pancreatic Neoplasms / genetics
  • Pancreatic Neoplasms / metabolism
  • Pancreatic Neoplasms / pathology
  • Patch-Clamp Techniques
  • Pyrimidinones / pharmacology
  • RNA Interference
  • TRPM Cation Channels / agonists
  • TRPM Cation Channels / antagonists & inhibitors
  • TRPM Cation Channels / metabolism*
  • Thiophenes / pharmacology

Substances

  • Benzamides
  • Membrane Proteins
  • N-(3-aminopropyl)-2-(((3-methylphenyl) methyl)oxy)-N-(2-thienylmethyl)benzamide hydrochloride salt
  • Pyrimidinones
  • TRPM Cation Channels
  • TRPM8 protein, human
  • Thiophenes
  • Menthol
  • icilin