A cell type-specific role of protein tyrosine phosphatase non-receptor type 2 in regulating ER stress signalling

Digestion. 2015;91(3):248-56. doi: 10.1159/000375459. Epub 2015 Mar 25.

Abstract

Background/aims: Genetic polymorphisms within the gene locus encoding protein tyrosine phosphatase non-receptor type 2 (PTPN2) have been associated with inflammatory bowel disease (IBD). A recent study demonstrated that PTPN2 regulates ER stress signalling in pancreatic β-cells. Therefore, we investigated whether PTPN2 regulates ER stress pathways, apoptosis and cytokine secretion in human intestinal epithelial cells (IECs) and monocytes.

Methods: THP-1 and HT-29 IECs were stimulated with 2 µg/ml tunicamycin (TNM) for the indicated periods of time. For knockdown experiments, cells were transfected using a mixture of three different PTPN2-specific siRNA oligonucleotides. Cell lysates were analysed by Western blot and real-time PCR. Cytokine secretion was studied by ELISA measurements of cell culture supernatant.

Results: TNM treatment reduced PTPN2 protein levels in HT-29 IECs and THP-1 monocytes. Knockdown of PTPN2 in THP-1 monocytes led to an exaggerated induction of phospho-eIF2α, enhanced PARP cleavage, indicative of apoptosis, and attenuated IL-8 and TNF secretion upon TNM stimulation. In HT-29 cells PTPN2 deficiency caused reduced phosphorylation of eIF2α and PARP cleavage under ER stress conditions.

Conclusions: Whereas the knockdown of PTPN2 made THP-1 cells more susceptible to ER stress, PTPN2 deficiency reduced ER stress responses in HT-29 IECs. This suggests that PTPN2 regulates adaptation to ER stress in a cell type-specific manner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Apoptosis / drug effects
  • Blotting, Western
  • Cytokines / metabolism
  • Endoplasmic Reticulum Stress / drug effects*
  • Endoplasmic Reticulum Stress / physiology
  • Epithelial Cells / metabolism*
  • Eukaryotic Initiation Factor-2 / metabolism
  • Gene Knockdown Techniques
  • HT29 Cells
  • Humans
  • Interleukin-8 / metabolism
  • Intestinal Mucosa / metabolism*
  • Monocytes / drug effects
  • Monocytes / metabolism
  • Phosphorylation
  • Protein Tyrosine Phosphatase, Non-Receptor Type 2 / genetics
  • Protein Tyrosine Phosphatase, Non-Receptor Type 2 / metabolism*
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • RNA, Small Interfering
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction / drug effects*
  • Tumor Necrosis Factor-alpha / metabolism
  • Tunicamycin / pharmacology

Substances

  • Anti-Bacterial Agents
  • Cytokines
  • Eukaryotic Initiation Factor-2
  • Interleukin-8
  • Proto-Oncogene Proteins
  • RNA, Small Interfering
  • Tumor Necrosis Factor-alpha
  • Tunicamycin
  • PTPN2 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 2