Effect of microRNA-27a-5p on apoptosis and inflammatory response of pancreatic acinar cells in acute pancreatitis by targeting PTEN

J Cell Biochem. 2019 Sep;120(9):15844-15850. doi: 10.1002/jcb.28855. Epub 2019 May 20.

Abstract

To investigate the apoptosis and inflammatory response of microRNA-27a-5p (miR-27a-5p) in pancreatic acinar cells of acute pancreatitis (AP) and its related mechanisms. Rat pancreatic acinar cell line AR42J was treated with caerulein (10nmol/L) to construct an acute pancreatitis cell model. Quantitative real-time polymerase chain reaction was performed to measure the expression of miR-27a-5p; The miR-27a-5p mimic was transfected into cell, and the apoptosis rate of the cells was detected by flow cytometry; The levels of TNF-α, IL-1, and IL-6 in the culture supernatant were determined by enzyme-linked immunosorbent assay; TargetScans database predicted and dual luciferase reporter gene assay verified the relationship between miR-27a-5p and the phosphatase and tensin homolog deleted on chromosome 10 (PTEN); The recovery experiment explored the apoptosis and the effects of inflammatory responses. The expression of miR-27a-5p decreased gradually (P < 0.05) and the expression of PTEN increased gradually (P < 0.05) with the prolongation of acting time. Upregulation of miR-27a-5p significantly promoted cell apoptosis (P < 0.05) and inhibited inflammatory response (P < 0.05); The TargetScans database predicted that the 3'UTR of PTEN contains a base complementary to the miR-27a-5p seed region. Cotransfection of wild-type vector (PTEN-WT) with miR-27a-5p mimic or miR-27a-5p inhibitor significantly affected the relative activity of luciferase (P < 0.05), and no significant impact was observed in mutant PTEN-MUT. Compared with miR-27a-5p + pcDNA group, transfection of miR-27a-5p mimic and pcDNA-PTEN significantly increased the expression of PTEN (P < 0.05), decreased the apoptotic rate (P < 0.05), and increased the inflammatory response (P < 0.05). miR-27a-5p induced apoptosis and inhibited the inflammatory response of pancreatic acinar cells in AP by targeting PTEN.

Keywords: acute pancreatitis; apoptosis; inflammation; miR-27a-5p.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acinar Cells / cytology*
  • Acinar Cells / drug effects
  • Acinar Cells / metabolism
  • Animals
  • Apoptosis
  • Cell Line
  • Cell Survival / drug effects
  • Ceruletide / adverse effects*
  • Gene Expression Regulation / drug effects
  • Interleukin-1 / metabolism
  • Interleukin-6 / metabolism
  • MicroRNAs / genetics*
  • PTEN Phosphohydrolase / genetics*
  • Pancreatitis / chemically induced
  • Pancreatitis / genetics*
  • Rats
  • Tumor Necrosis Factor-alpha / metabolism
  • Up-Regulation

Substances

  • Il6 protein, rat
  • Interleukin-1
  • Interleukin-6
  • MIRN27 microRNA, rat
  • MicroRNAs
  • Tumor Necrosis Factor-alpha
  • Ceruletide
  • PTEN Phosphohydrolase
  • Pten protein, rat