Fluctuation of ROS regulates proliferation and mediates inhibition of migration by reducing the interaction between DLC1 and CAV-1 in breast cancer cells

In Vitro Cell Dev Biol Anim. 2017 Apr;53(4):354-362. doi: 10.1007/s11626-016-0123-0. Epub 2017 Jan 27.

Abstract

The aim of our present study was to elucidate the effects of up-regulation and down-regulation of intracellular reactive oxygen species (ROS) level on proliferation, migration, and related molecular mechanism. Breast cancer cells were treated by catalase or H2O2. MTT, colony formation assay, and Hoechst/PI staining were used to evaluate proliferation and apoptosis. The level of intracellular ROS was measured by dichlorodihydrofluorescein diacetate probes. The ability of migration was detected by wound healing. Western blotting and coimmunoprecipitation (co-IP) were used to determine the expression of DLC1 and CAV-1 and their interaction. Our data indicated that up-regulation of intracellular ROS induced by H2O2 significantly inhibited proliferation and induced apoptosis accompanying G1 cell cycle arrest and elevated expression of p53. For cell migration, either up-regulation or down-regulation of ROS induced migration inhibition with reduction of interaction between DLC1 and CAV-1. Our results suggested that up-regulation of intracellular ROS inhibited proliferation by promoting expression of p53 and induced G1 cycle arrest and apoptosis. Fluctuation of ROS inhibited migration through reducing the interaction between DLC1 and CAV-1.

Keywords: CAV-1; DLC1; Migration; Proliferation; ROS.

MeSH terms

  • Apoptosis
  • Breast Neoplasms / metabolism*
  • Breast Neoplasms / pathology*
  • Caveolin 1 / metabolism*
  • Cell Cycle Checkpoints
  • Cell Line, Tumor
  • Cell Movement*
  • Cell Proliferation
  • Cell Survival
  • Female
  • GTPase-Activating Proteins / metabolism*
  • Humans
  • Hydrogen Peroxide / metabolism
  • Protein Binding
  • Reactive Oxygen Species / metabolism*
  • Tumor Suppressor Protein p53 / metabolism
  • Tumor Suppressor Proteins / metabolism*

Substances

  • Caveolin 1
  • DLC1 protein, human
  • GTPase-Activating Proteins
  • Reactive Oxygen Species
  • Tumor Suppressor Protein p53
  • Tumor Suppressor Proteins
  • Hydrogen Peroxide