Novel miR-29b target regulation patterns are revealed in two different cell lines

Sci Rep. 2019 Nov 25;9(1):17449. doi: 10.1038/s41598-019-53868-x.

Abstract

MicroRNAs (miRNAs) are a class of small non-coding RNAs that regulate gene or protein expression by targeting mRNAs and triggering either translational repression or mRNA degradation. Distinct expression levels of miRNAs, including miR-29b, have been detected in various biological fluids and tissues from a large variety of disease models. However, how miRNAs "react" and function in different cellular environments is still largely unknown. In this study, the regulation patterns of miR-29b between human and mouse cell lines were compared for the first time. CRISPR/Cas9 gene editing was used to stably knockdown miR-29b in human cancer HeLa cells and mouse fibroblast NIH/3T3 cells with minimum off-targets. Genome editing revealed mir-29b-1, other than mir-29b-2, to be the main source of generating mature miR-29b. The editing of miR-29b decreased expression levels of its family members miR-29a/c via changing the tertiary structures of surrounding nucleotides. Comparing transcriptome profiles of human and mouse cell lines, miR-29b displayed common regulation pathways involving distinct downstream targets in macromolecular complex assembly, cell cycle regulation, and Wnt and PI3K-Akt signalling pathways; miR-29b also demonstrated specific functions reflecting cell characteristics, including fibrosis and neuronal regulations in NIH/3T3 cells and tumorigenesis and cellular senescence in HeLa cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • CRISPR-Cas Systems
  • Cell Transformation, Neoplastic
  • Cellular Senescence
  • Clone Cells
  • Gene Editing
  • Gene Expression Regulation*
  • Gene Knockdown Techniques
  • Gene Ontology
  • HeLa Cells / metabolism*
  • Humans
  • Mice
  • MicroRNAs / biosynthesis
  • MicroRNAs / genetics*
  • Models, Molecular
  • NIH 3T3 Cells / metabolism*
  • Nucleic Acid Conformation
  • RNA, Guide / genetics
  • Sequence Alignment
  • Signal Transduction
  • Transcriptome

Substances

  • MIRN29 microRNA, human
  • MIRN29 microRNA, mouse
  • MicroRNAs
  • RNA, Guide