GSK3β negatively regulates HIF1α mRNA stability via nucleolin in the MG63 osteosarcoma cell line

Biochem Biophys Res Commun. 2014 Jan 10;443(2):598-603. doi: 10.1016/j.bbrc.2013.12.020. Epub 2013 Dec 11.

Abstract

Hypoxia-inducible factor 1α (HIF1α) is a transcription factor involved in the growth, invasion and metastasis of malignant tumors. Glycogen synthase kinase 3 beta (GSK3β) is a protein kinase involved in a variety of signaling pathways, such as the Wnt and NF-κB pathways; this kinase can affect tumor progress through the regulation of transcription factor expression and apoptosis. Recent studies showed that GSK3β was involved in the expression of HIF1α. However, the effect of GSK3β on HIF1α expression in osteosarcoma cells remains unknown. To understand the relationship between GSK3β and HIF1α comprehensively, small RNA interference techniques, Western blot analyses, quantitative real-time PCR analyses and luciferase assays were used in our study. Experimental data revealed that inhibition of GSK3β could increase HIF1α protein levels and expression of its target genes by increasing the stability of the HIF1α mRNA, not by affecting the HIF1α protein stability, and that this process could be mediated by nucleolin.

Keywords: ARNT; Chx; EPO; GLUT1; GSK3β; HIF1α; Nucleolin; ODD; Osteosarcoma; SDS–PAGE; VEGF; VHL; aryl hydrocarbon receptor nuclear translocator; cycloheximide; erythropoietin; glucose transporter 1; glycogen synthase kinase 3 beta; hypoxia-inducible factor 1α; mRNA; oxygen-dependent degradation; siRNA; small interfering RNA; sodium dodecyl sulfate polyacrylamide gel electrophoresis; vascular endothelial growth factor; von Hippel-Lindau.

MeSH terms

  • Cell Line, Tumor
  • Down-Regulation
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • Nucleolin
  • Osteosarcoma / metabolism*
  • Phosphoproteins / metabolism*
  • RNA Stability / physiology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • RNA-Binding Proteins / metabolism*

Substances

  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Phosphoproteins
  • RNA, Messenger
  • RNA-Binding Proteins