MicroRNA-7a inhibits Isl1 expression to regulate insulin secretion by targeting Raf1 and Mapkap1 in NIT-1 cells

In Vitro Cell Dev Biol Anim. 2021 Sep;57(8):817-824. doi: 10.1007/s11626-021-00611-4. Epub 2021 Oct 28.

Abstract

Both microRNA-7a (miR-7a) and LIM-homeodomain transcription factor ISL1 are important factors regulating insulin transcription and secretion, but the functional relationship and the interacting mechanisms between miR-7a and ISL1 in pancreatic islet β-cells remain unknown. The aims of this study were thus to identify the potential interactions and signaling communication between miR-7a and ISL1 in regulating insulin transcription and secretion in the cultured NIT-1 cells. The results show that miR-7a inhibitor upregulates Isl-1 and insulin gene expressions, and the insulin secretion. Whereas miR-7a mimics inhibit ISL1 and insulin gene expressions, and decreases the insulin secretion. Furthermore, we identified the target gene of miR-7a using dual-luciferase reporter assay, and the results demonstrate that Raf1 and Mapkap1 is a direct target gene of miR-7a, modeling RAF1/MEK/ERK1/2 and mTORC2/AKT signaling pathway to regulate Isl1 expression, and thus influencing insulin expression and secretion. Our results indicate that therapeutic inhibition of miR-7a function could be of relevance for preserving the function of pancreatic β-cells during the course of diabetes development, implicating miR-7, ISL1, and/or the connecting molecules may act as novel targets for pharmacological or gene therapy in diabetes and related metabolic disease, although much detailed studies are required in the further study.

Keywords: ISL1; Insulin; Mapkap1; Raf1; miR-7a.

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Line
  • Insulin / metabolism
  • Insulin Secretion* / physiology
  • Insulin-Secreting Cells / metabolism*
  • Insulin-Secreting Cells / physiology
  • LIM-Homeodomain Proteins / metabolism*
  • Mice
  • MicroRNAs / metabolism*
  • Proto-Oncogene Proteins c-raf / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Ribosomal Protein S6 Kinases, 90-kDa / metabolism*
  • Transcription Factors / metabolism*

Substances

  • Insulin
  • LIM-Homeodomain Proteins
  • MIRN7 microRNA, mouse
  • MicroRNAs
  • Transcription Factors
  • insulin gene enhancer binding protein Isl-1
  • Proto-Oncogene Proteins c-raf
  • Raf1 protein, mouse
  • Ribosomal Protein S6 Kinases, 90-kDa