miR-106b is overexpressed in medulloblastomas and interacts directly with PTEN

Neuropathol Appl Neurobiol. 2015 Feb;41(2):145-64. doi: 10.1111/nan.12169.

Abstract

Aims: MicroRNAs (miRNAs) are an abundant group of small non-coding RNAs that have been implicated in tumorigenesis. They regulate expression of target genes by complementary base pairing. The purposes of this study were to delineate miR-106b expression in medulloblastoma (MB) and to explore its functional contributions to MB pathogenesis.

Methods: We analysed expression of miR-106b in 32 MB samples by quantitative RT-PCR. We applied gain- and loss-of-function strategies to delineate the functional roles of miR-106b in MB. Luciferase reporter assay was conducted to confirm target gene of miR-106b.

Results: Expression of miR-106b was overexpressed in MB, and was significantly associated with its host gene MCM7 (P = 0.020). Transfection of miR-106b inhibitor in MB cell lines markedly reduced cell proliferation, migration and invasion potential, and tumour sphere formation. Cell cycle analysis indicated that miR-106b inhibition induced G1 arrest and apoptosis. The cell cycle regulators, p21 and cyclin D1, and apoptotic marker cleaved PARP were differentially expressed in miR-106b inhibitor-transfected cells. PTEN was identified as a direct target gene of miR-106b. Luciferase reporter assay confirmed miR-106b directly interacted with the 3' UTR of PTEN. We found miR-106b directly targeted PTEN at transcriptional and translational levels. Immunohistochemistry revealed a trend between PTEN and miR-106b in MB tumours (P = 0.07).

Conclusions: These data suggested the upregulation of miR-106b in MB and the involvement of miR-106b in MB biology.

Keywords: MCM7; PTEN; medulloblastoma; miR-106b; microRNA.

MeSH terms

  • Adolescent
  • Adult
  • Blotting, Western
  • Cell Line, Tumor
  • Cerebellar Neoplasms / genetics*
  • Cerebellar Neoplasms / metabolism
  • Child
  • Female
  • Gene Expression Regulation, Neoplastic / genetics*
  • Humans
  • Immunohistochemistry
  • Male
  • Medulloblastoma / genetics*
  • Medulloblastoma / metabolism
  • MicroRNAs / analysis
  • MicroRNAs / biosynthesis*
  • PTEN Phosphohydrolase / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Transfection
  • Up-Regulation
  • Young Adult

Substances

  • MIRN106 microRNA, human
  • MicroRNAs
  • PTEN Phosphohydrolase
  • PTEN protein, human