Aristolochic acid inhibits Slit2-induced migration and tube formation via inactivation of Robo1/Robo2-NCK1/NCK2 signaling pathway in human umbilical vein endothelial cells

Toxicol Lett. 2019 Jan:300:51-58. doi: 10.1016/j.toxlet.2018.10.022. Epub 2018 Oct 28.

Abstract

Robo1/Robo2-NCK1/NCK2 signaling pathway controls endothelial cell sprouting and migration induced by Slit2 or VEGF, but whether it is involved in peritubular capillary (PTC) rarefaction of Aristolochic acid nephropathy (AAN) is unclear. In the present study, we evaluated whether AA exerts antiangiogenic effects by targeting this signaling pathways in HUVECs. HUVECs or lentivirus-mediated NCK1-overexpressing HUVECs were stimulated with AA (1, 2 or 3 μg/ml) in the absence or presence of 6 nM Slit2. Our results showed that AAІ (1-3 μg/ml) dose-dependently inhibited the migration and tube formation of HUVECs. This inhibition was in parallel with down-regulated mRNA and protein expression of Slit2/Robo1/Robo2-NCK1/NCK2 signaling pathway. Importantly, overexpression of NCK1 rescued AAІ-impaired angiogenesis, as evidenced by the increase of cell migration and tube formation of HUVECs in response to Slit2. The down-regulation of NCK2 and decreased activation of Rac1 was also restored by overexpression of NCK1. Taken together, our findings show that AA inhibits Slit2-induced migration and tube formation via inactivation of Robo1/Robo2-NCK1/NCK2 signaling pathway in HUVECs, and NCK1 might be a potential agent for vascular remodeling in AAN and diseases associated with impaired angiogenesis.

Keywords: Angiogenesis; Aristolochic acid nephropathy; Endothelial cells; NCK; Rac1; Slit2/Robo1 signaling.

MeSH terms

  • Angiogenesis Inhibitors / metabolism
  • Aristolochic Acids / toxicity*
  • Carcinogens / toxicity*
  • Cell Movement / drug effects*
  • Cell Proliferation / drug effects*
  • Human Umbilical Vein Endothelial Cells / drug effects*
  • Humans
  • Microvascular Rarefaction / chemically induced*
  • Receptors, Immunologic / metabolism
  • Signal Transduction / drug effects*

Substances

  • Angiogenesis Inhibitors
  • Aristolochic Acids
  • Carcinogens
  • ROBO2 protein, human
  • Receptors, Immunologic