miR-198 Represses the Proliferation of HaCaT Cells by Targeting Cyclin D2

Int J Mol Sci. 2015 Jul 27;16(8):17018-28. doi: 10.3390/ijms160817018.

Abstract

Background: MiR-198 has been considered as an inhibitor of cell proliferation, invasion, migration and a promoter of apoptosis in most cancer cells, while its effect on non-cancer cells is poorly understood.

Methods: The effect of miR-198 transfection on HaCaT cell proliferation was firstly detected using Cell Count Kit-8 and the cell cycle progression was analyzed by flow cytometry. Using bioinformatics analyses and luciferase assay, a new target of miR-198 was searched and identified. Then, the effect of the new target gene of miR-198 on cell proliferation and cell cycle was also detected.

Results: Here we showed that miR-198 directly bound to the 3'-UTR of CCND2 mRNA, which was a key regulator in cell cycle progression. Overexpressed miR-198 repressed CCND2 expression at mRNA and protein levels and subsequently led to cell proliferation inhibition and cell cycle arrest in the G1 phase. Transfection ofSiCCND2 in HaCaT cells showed similar inhibitory effects on cell proliferation and cell cycle progression.

Conclusion: In conclusion, we have identified that miR-198 inhibited HaCaT cell proliferation by directly targeting CCND2.

Keywords: HaCaT; cell proliferation; cyclin D2; miR-198.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics
  • Base Sequence
  • Binding Sites
  • Cell Cycle Checkpoints / genetics
  • Cell Line
  • Cell Proliferation
  • Cyclin D2 / genetics*
  • Cyclin D2 / metabolism
  • Enzyme Assays
  • G1 Phase / genetics
  • Humans
  • Keratinocytes / cytology*
  • Keratinocytes / metabolism*
  • Luciferases / metabolism
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Molecular Sequence Data
  • Protein Binding / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Transfection

Substances

  • 3' Untranslated Regions
  • CCND2 protein, human
  • Cyclin D2
  • MIRN198 microRNA, human
  • MicroRNAs
  • RNA, Messenger
  • RNA, Small Interfering
  • Luciferases