miR-15b reduces amyloid-β accumulation in SH-SY5Y cell line through targetting NF-κB signaling and BACE1

Biosci Rep. 2018 Nov 13;38(6):BSR20180051. doi: 10.1042/BSR20180051. Print 2018 Dec 21.

Abstract

Alzheimer's disease (AD) is the multifactorial neurodegenerative disorder causing progressive memory loss and cognitive impairment. The aberrant accumulation of amyloid-β (Aβ) and neuroinflammation are two major events in AD. BACE1 is required for the cleavage of amyloid precursor protein (APP) to generate Aβ, which stimulates the nuclear transcription factor κB (NF-κB) signaling, leading to the secretion of inflammatory cytokines. And NF-κB can up-regulate the expression of BACE1. miRNAs are small non-coding RNAs that regulate gene transcription. miR-15b down-regulates BACE1 expression while it is unclear whether miR-15b can regulate Aβ in human neuronal cells, and if so, whether it is by targetting NF-κB. SH-SY5Y cell line was transfected with Swedish APP mutant (APPswe) as an in vitro AD model. Quantitative PCR (qPCR), WB, and ELISA were used to detected related gene expression intracellularly or in supernatant. Dual luciferase assay was used to validate miRNA and targets binding. miR-15b inhibits expression of BACE1and APP. Moreover, the reduced level of Aβ was observed in response to miR-15b mimics in SH-SH5Y/APPswe cells. miR-15b directly targetted the conserved Bace1 3'UTR to regulate its expression. In addition, the inhibition of APPswe-induced secretion of inflammatory cytokines and the suppression of NF-κB activation by miR-15b were validated. And miR-15b directly targetted the 3'UTRs of NF-κB1 and inhibitor of NF-κB (IκB) kinase α (IKK-α), encoding NF-κB1 and IKK-α, respectively. Our study suggests that miR-15b inhibits Aβ accumulation through targetting NF-κB signaling and BACE1 and serves as a potential molecular target for AD therapy.

Keywords: Alzheimer’s disease; Aβ; BACE1; NF-κB; miR-15b.

MeSH terms

  • 3' Untranslated Regions / genetics
  • Alzheimer Disease / genetics*
  • Alzheimer Disease / pathology
  • Amyloid Precursor Protein Secretases / genetics*
  • Amyloid beta-Peptides / genetics
  • Amyloid beta-Peptides / metabolism
  • Amyloid beta-Protein Precursor / genetics*
  • Aspartic Acid Endopeptidases / genetics*
  • Gene Expression Regulation
  • Humans
  • I-kappa B Kinase / genetics
  • MicroRNAs / genetics*
  • Mutation
  • NF-kappa B / genetics
  • NF-kappa B p50 Subunit / genetics
  • Neurons / metabolism
  • Neurons / pathology
  • Protein Aggregation, Pathological / genetics
  • Signal Transduction / genetics
  • Transcription Factor RelA / genetics

Substances

  • 3' Untranslated Regions
  • Amyloid beta-Peptides
  • Amyloid beta-Protein Precursor
  • MIRN15 microRNA, human
  • MicroRNAs
  • NF-kappa B
  • NF-kappa B p50 Subunit
  • NFKB1 protein, human
  • RELA protein, human
  • Transcription Factor RelA
  • I-kappa B Kinase
  • Amyloid Precursor Protein Secretases
  • Aspartic Acid Endopeptidases
  • BACE1 protein, human