Up-regulated MiR-27-3p promotes the G1-S phase transition by targeting inhibitor of growth family member 5 in osteosarcoma

Biomed Pharmacother. 2018 May:101:219-227. doi: 10.1016/j.biopha.2018.02.066. Epub 2018 Feb 26.

Abstract

Objective: MicroRNAs (miRNAs) play an essential role in regulating malignant progression of tumour cells by inhibiting translation or stability of messenger RNA. However, the expression pattern and regulatory mechanism of miR-27-3p in osteosarcoma remains unclear.

Methods: We examined the expression of miR-27-3p in 5 osteosarcoma cell lines compared with that in 2 normal osteocyte cell lines. Osteosarcoma cells U-2OS and MG-63 were transduced to up-regulate or down-regulate the expression of miR-27-3p. The 3-(4, 5-Dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide, or MTT, assay, colony formation assays, BrdUrd labelling, immunofluorescence, anchorage-independent growth ability assay and flow cytometry analysis were used to test the effect of miR-27-3p. Luciferase assays were added to verify the direct relationship between miR-27-3p and the predicted target gene inhibitor of growth family member 5 (ING5).

Results: The expression of miR-27-3p was significantly increased in examined osteosarcoma cell lines compared with that in normal osteocyte cell lines. Up-regulation of miR-27-3p significantly accelerated osteosarcoma cell growth via promoting G1-S transition. In addition, the opposite result was observed in miR-27-3p-down-regulated cells. Up-regulation of ING5 significantly attenuated the miR-27-3p-induced proliferation in osteosarcoma cells.

Conclusions: These data suggested that miR-27-3p could promote the G1-S phase transition that leads to proliferation by down-regulating the expression of ING5 in osteosarcoma.

Keywords: G1-S phase; ING5; Osteosarcoma; Proliferation; miR-27-3p.

MeSH terms

  • Bone Neoplasms / genetics*
  • Bone Neoplasms / pathology
  • Cell Line
  • Cell Line, Tumor
  • Cell Proliferation / genetics
  • Down-Regulation
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • G1 Phase / genetics
  • Gene Expression Regulation, Neoplastic
  • Humans
  • MicroRNAs / genetics*
  • Osteocytes / metabolism
  • Osteosarcoma / genetics*
  • Osteosarcoma / pathology
  • S Phase / genetics
  • Transcription Factors / genetics*
  • Tumor Suppressor Proteins / genetics*
  • Up-Regulation

Substances

  • ING5 protein, human
  • MIRN27 microRNA, human
  • MicroRNAs
  • Transcription Factors
  • Tumor Suppressor Proteins