Differential transcriptional regulation of hypoxia-inducible factor-1α by arsenite under normoxia and hypoxia: involvement of Nrf2

J Mol Med (Berl). 2016 Oct;94(10):1153-1166. doi: 10.1007/s00109-016-1439-7. Epub 2016 Jun 10.

Abstract

Arsenite (As(III)) is widely distributed in nature and can be found in water, food, and air. There is significant evidence that exposure to As(III) is associated with human cancers originated from liver, lung, skin, bladder, kidney, and prostate. Hypoxia plays a role in tumor growth and aggressiveness; adaptation to it is, at least to a large extent, mediated by hypoxia-inducible factor-1α (HIF-1α). In the current study, we investigated As(III) effects on HIF-1α under normoxia and hypoxia in the hepatoma cell line HepG2. We found that As(III) increased HIF-1α protein levels under normoxia while the hypoxia-mediated induction of HIF1α was reduced. Thereby, the As(III) effects on HIF-1α were dependent on both, transcriptional regulation via the transcription factor Nrf2 mediated by NOX4, PI3K/Akt, and ERK1/2 as well as by modulation of HIF-1α protein stability. In line, the different effects of As(III) via participation of HIF-1α and Nrf2 were also seen in tube formation assays with endothelial cells where knockdown of Nrf2 and HIF-1α abolished As(III) effects. Overall, the present study shows that As(III) is a potent inducer of HIF-1α under normoxia but not under hypoxia which may explain, in part, its carcinogenic as well as anti-carcinogenic actions.

Key message: As(III) increased HIF-1α under normoxia but reduced its hypoxia-dependent induction. The As(III) effects on HIF-1α were dependent on ROS, NOX4, PI3K/Akt, and ERK1/2. The As(III) effects under normoxia involved transcriptional regulation via Nrf2. Knockdown of Nrf2 and HIF-1α abolished As(III) effects in tube formation assays. The data may partially explain As(III)'s carcinogenic and anti-carcinogenic actions.

Keywords: Arsenite As(III); Hypoxia-inducible factor 1 (HIF-1α); Mitogen-activated protein kinase (MAPK); NADPH enzyme oxidase 4 (NOX4); Reactive oxygen species (ROS).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Arsenites / pharmacology*
  • Carcinogens / pharmacology
  • Cell Hypoxia / genetics*
  • Cell Hypoxia / physiology
  • Cell Line
  • Cells, Cultured
  • Extracellular Signal-Regulated MAP Kinases
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Gene Expression Regulation / drug effects*
  • Heme Oxygenase-1 / genetics
  • Hep G2 Cells
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Mice
  • NADPH Oxidase 4
  • NADPH Oxidases
  • NF-E2-Related Factor 2
  • Phosphatidylinositol 3-Kinases
  • Plasminogen Activator Inhibitor 1 / genetics
  • Proto-Oncogene Proteins c-akt
  • Reactive Oxygen Species / metabolism

Substances

  • Antineoplastic Agents
  • Arsenites
  • Carcinogens
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • NF-E2-Related Factor 2
  • NFE2L2 protein, human
  • Plasminogen Activator Inhibitor 1
  • Reactive Oxygen Species
  • Heme Oxygenase-1
  • NADPH Oxidase 4
  • NADPH Oxidases
  • NOX4 protein, human
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • Extracellular Signal-Regulated MAP Kinases
  • arsenite