Silencing Drp1 inhibits glioma cells proliferation and invasion by RHOA/ ROCK1 pathway

Biochem Biophys Res Commun. 2016 Sep 16;478(2):663-8. doi: 10.1016/j.bbrc.2016.08.003. Epub 2016 Aug 3.

Abstract

Backgrounds: Dynamin-related protein 1 (Drp1) is a newly discovered therapeutic target for tumor initiation, migration, proliferation, and chemosensitivity. In the present study, we aimed to examine the level of expression and distribution of DRP1 in glioma tissues and explore the concrete mechanism of DRP1 played in glioma.

Methods: Expression of DRP1 in glioma tissues was determined by immunohistochemistry staining. The DRP1 gene was knocked down using small interfering RNA, and was overexpressed using plasmids in glioma cells. To assess changes in cell function, in vitro assays for invasion and growth were applied. Protein expression was tested by using Western-blot method. Variation of F-actin in cells was analyzed using immunofluorescence staining. Interactions between proteins were determined by co-immunoprecipitation.

Results: The protein expression levels of DRP1 were significantly increased in glioma tissues compared to the normal brain tissues. Down-regulation of DRP1 decreased cell proliferation and invasion, and inhibited the formation of pseudopodias and microvillis. Moreover, a possible link between DRP1 and RHOA was confirmed when interactions between these two proteins were observed in the cells.

Conclusions: Our results demonstrated that silencing DRP1 regulated the cytoskeleton remodeling through inhibiting RHOA/ROCK1 pathway, and thus decreased the proliferation and invasion of glioma cells.

Keywords: DRP1; Glioma; RHOA; ROCK1.

MeSH terms

  • Actins / genetics
  • Actins / metabolism
  • Brain Neoplasms / genetics*
  • Brain Neoplasms / metabolism
  • Brain Neoplasms / pathology
  • Case-Control Studies
  • Cell Line, Tumor
  • Cell Movement
  • Cell Proliferation
  • Cytoskeleton / metabolism
  • Cytoskeleton / ultrastructure
  • Dynamins
  • GTP Phosphohydrolases / antagonists & inhibitors
  • GTP Phosphohydrolases / genetics*
  • GTP Phosphohydrolases / metabolism
  • Gene Expression Regulation, Neoplastic*
  • Gene Silencing*
  • Glioma / genetics*
  • Glioma / metabolism
  • Glioma / pathology
  • Humans
  • Microtubule-Associated Proteins / antagonists & inhibitors
  • Microtubule-Associated Proteins / genetics*
  • Microtubule-Associated Proteins / metabolism
  • Microvilli / metabolism
  • Microvilli / ultrastructure
  • Mitochondrial Proteins / antagonists & inhibitors
  • Mitochondrial Proteins / genetics*
  • Mitochondrial Proteins / metabolism
  • Pseudopodia / metabolism
  • Pseudopodia / ultrastructure
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Signal Transduction
  • Tissue Array Analysis
  • rho-Associated Kinases / genetics*
  • rho-Associated Kinases / metabolism
  • rhoA GTP-Binding Protein / genetics*
  • rhoA GTP-Binding Protein / metabolism

Substances

  • Actins
  • Microtubule-Associated Proteins
  • Mitochondrial Proteins
  • RNA, Small Interfering
  • RHOA protein, human
  • ROCK1 protein, human
  • rho-Associated Kinases
  • GTP Phosphohydrolases
  • rhoA GTP-Binding Protein
  • DNM1L protein, human
  • Dynamins