Downregulation of MiR-93 expression reduces cell proliferation and clonogenicity of HepG2 cells

Hepatogastroenterology. 2012 Nov-Dec;59(120):2367-73. doi: 10.5754/hge12458.

Abstract

Background/aims: MiR-93 was observed in various types of cancers. This study is to investigate a role of miR-93 in the carcinogenesis of HCC.

Methodology: The expression of miR-93 in HepG2 cells and prima-ry human hepatocytes (PHHC) was measured by RT-PCR. HepG2 cells were transfected with miR-93 inhibitor or negative control. The cell proliferation was determined by using the CellTiter 96® Aqueous One Solution Cell Proliferation Assay kit. The migration and clonogenicity in vitro were measured by cell migration assay, colony formation analysis and anchorage-in-dependent growth assay. The apoptosis and cell cycle were detected by flow cytometry analysis. The mRNA and protein levels of transforming growth factor-beta type II receptor (TGFBR2) and integrin beta8 (ITGB8)were evaluated by RT-PCR and western blot analysis.

Results: MiR-93 was upregulated in HepG2 cells compared with PHHC and inhibition of miR-93 significantly suppressed HepG2 cell proliferation, migration and col-ony formation. The expressions of TGFBR2 and ITGB8 were upregulated when miR-93 was inhibited.

Conclusions: Our results reveal an important contribution for miR-93 in hepatocarcinogenesis and suggest a role for TGFBR2 and ITGB8 dysregulation in this process. Thus,the use of synthetic inhibitor of miR-93 may prove to bea promising approach to liver cancer treatment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / genetics
  • Blotting, Western
  • Carcinoma, Hepatocellular / genetics*
  • Carcinoma, Hepatocellular / metabolism
  • Carcinoma, Hepatocellular / pathology
  • Cell Adhesion / genetics
  • Cell Cycle Checkpoints / genetics
  • Cell Movement / genetics
  • Cell Proliferation*
  • Down-Regulation
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic
  • Hep G2 Cells
  • Humans
  • Integrin beta Chains / genetics
  • Integrin beta Chains / metabolism
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology
  • MicroRNAs / metabolism*
  • Primary Cell Culture
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Receptor, Transforming Growth Factor-beta Type II
  • Receptors, Transforming Growth Factor beta / genetics
  • Receptors, Transforming Growth Factor beta / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection
  • Tumor Stem Cell Assay

Substances

  • Integrin beta Chains
  • MIRN93 microRNA, human
  • MicroRNAs
  • RNA, Messenger
  • Receptors, Transforming Growth Factor beta
  • integrin beta8
  • Protein Serine-Threonine Kinases
  • Receptor, Transforming Growth Factor-beta Type II