Apoptosis Induced by (+)-Betulin Through NF-κB Inhibition in MDA-MB-231 Breast Cancer Cells

Anticancer Res. 2020 Dec;40(12):6637-6647. doi: 10.21873/anticanres.14688. Epub 2020 Dec 7.

Abstract

Background/aim: This study aimed to uncover the effects of (+)-betulin on the NF-κB-apoptotic pathway in MDA-MB-231 cells, and determine its toxicity and protein expression in vivo.

Materials and methods: Cell cytotoxicity and toxicity were determined using the SRB assay and a zebrafish model, respectively. Western blot, mitochondrial transmembrane potential (MTP), and computational modeling analysis were performed.

Results: (+)-betulin inhibited the growth of MDA-MB-231 cells, but did not induce toxicity in zebrafish. (+)-Betulin inhibited the activity of NF-κB p65 in silico and in vitro. In cells, (+)-betulin down-regulated NF-κB p50 and 65, IKKα and β, ICAM-1 and bcl-2 expressions. Cell co-treatment with (+)-betulin and TNFα increased the (+)-betulin cytotoxic potential. Moreover, (+)-betulin induced the loss of MTP. Furthermore, (+)-betulin, in zebrafish, down-regulated the expression of NF-κB p65, IKKα, ΙΚΚβ and procaspase-3.

Conclusion: The results contribute to the understanding of the mode of action on apoptosis induction by inhibiting NF-κB pathway in MDA-MB-231 cells.

Keywords: (+)-betulin; MDA-MB-231; NF-κB; apoptosis; zebrafish.

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Breast Neoplasms / pathology*
  • Cell Line, Tumor
  • Female
  • Humans
  • Membrane Potential, Mitochondrial / drug effects
  • Molecular Docking Simulation
  • NF-kappa B / antagonists & inhibitors*
  • NF-kappa B / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Signal Transduction / drug effects
  • Triterpenes / chemistry
  • Triterpenes / pharmacology*
  • Tumor Necrosis Factor-alpha / pharmacology
  • Zebrafish

Substances

  • NF-kappa B
  • Proto-Oncogene Proteins c-bcl-2
  • Triterpenes
  • Tumor Necrosis Factor-alpha
  • betulin