Altered Biological Properties in Dp71 Over-Expressing HBE Cells

Cell Physiol Biochem. 2017;43(5):2022-2036. doi: 10.1159/000484181. Epub 2017 Oct 23.

Abstract

Background/aims: In order to further characterize the biological traits of Dp71, HBE over expressing two most abundantly expressed Dp71 spliced isoforms, Dp71d and Dp71f, were established and their biological traits were explored.

Methods: The proliferation, migration and invasion capabilities of HBE-Dp71d and HBE-Dp71f cells were evaluated by MTT, colony formation, transwell and scratch assay. Cell cycle and apoptosis induced by H2O2 were measured by flow cytometer. Co-IP was performed to prove the interaction between lamin B1, FAK and Dp71. Western blot was performed to detect lamin B1, FAK, ERK and Cyclin D expression in HBE-Dp71d and HBE-Dp71f cells.

Results: HBE-Dp71d and HBE-Dp71f cells proliferated faster than their mock and blank controls; shortened their G0/G1 phase; enhanced their invasion and migration capabilities; reduced their apoptosis induced by H2O2. Co-IP proved Dp71 directly interacting with focal adhesion kinase (FAK) and lamin B1 in HBE cells. Increased lamin B1, FAK mRNA and protein expression, over activation of integrin/focal adhesion kinase/extracellular signal-regulated kinase (ERK)/cyclin D pathway were observed in HBE-Dp71d and HBE-Dp71f cells.

Conclusions: Via increasing FAK in the cytoplasmic FAK-Dp71 , lamin B1 of nucleus laminB1-Dp71 complex, HBE-Dp71d and HBE-Dp71f cells alter their proliferation, migration, invasion, cell cycle and apoptosis rate induced by H2O2.

Keywords: Biological characters; Dp71; FAK /ERK/cyclin D; HBE; Lamin B1.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Cell Line
  • Cell Movement / drug effects
  • Cell Movement / genetics
  • Cell Proliferation / drug effects
  • Cell Proliferation / genetics
  • Cyclin D / metabolism
  • Dystrophin / genetics
  • Dystrophin / metabolism*
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Focal Adhesion Kinase 1 / metabolism
  • G1 Phase / genetics
  • G1 Phase / physiology
  • HeLa Cells
  • Humans
  • Hydrogen Peroxide / pharmacology
  • PC12 Cells
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Rats
  • Resting Phase, Cell Cycle / genetics
  • Resting Phase, Cell Cycle / physiology

Substances

  • Cyclin D
  • Dystrophin
  • Protein Isoforms
  • apo-dystrophin 1
  • Hydrogen Peroxide
  • Focal Adhesion Kinase 1
  • Extracellular Signal-Regulated MAP Kinases