ROS -mediated p53 activation by juglone enhances apoptosis and autophagy in vivo and in vitro

Toxicol Appl Pharmacol. 2019 Sep 15:379:114647. doi: 10.1016/j.taap.2019.114647. Epub 2019 Jul 5.

Abstract

Juglone (JG) exhibits a broad-spectrum of cytotoxicity against some cancer cells. However, its molecular mechanisms have not been investigated well. Here, the present results showed that JG significantly inhibited tumor growth in vivo. CCK-8 assays, flow cytometric analysis, western blotting and immunohistochemistry revealed that JG effectively inhibited cell proliferation and induced apoptosis through extrinsic pathways. We also observed that JG treatment induced autophagy flux via activiting the AMPK-mTOR signaling pathway. In addition, we found that JG enhanced p53 activation by increasing down-regulation of ubiquitin-mediated degradation. Inhibition of p53 by siRNA attenuated JG-induced cell death and autophagy. Moreover, JG enhanced the generation of hydrogen peroxide (H2O2) and superoxide anion radical (O2• -). Further experiments proved that H2O2 was a major factor since the H2O2 scavenger catalase (CAT) reduced both autophagy and cell death to a greater extent than the O2• - scavenger SOD. Overall, our results illustrated that JG caused apoptosis and autophagy via activating the ROS-mediated p53 pathway in human liver cancer cells in vitro and in vivo, which provided basic scientific evidence that JG serves as a potential anti-cancer agent.

Keywords: Apoptosis; Autophagy; HepG2 cells; Juglone; Reactive oxygen species; p53.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Autophagy / drug effects*
  • Flow Cytometry
  • Gene Expression / drug effects
  • Hep G2 Cells / drug effects
  • Hep G2 Cells / metabolism
  • Humans
  • Naphthoquinones / pharmacology*
  • Polymerase Chain Reaction
  • Reactive Oxygen Species / metabolism*
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • Naphthoquinones
  • Reactive Oxygen Species
  • Tumor Suppressor Protein p53
  • juglone