Impact of p53 status on TRAIL-mediated apoptotic and non-apoptotic signaling in cancer cells

PLoS One. 2019 Apr 4;14(4):e0214847. doi: 10.1371/journal.pone.0214847. eCollection 2019.

Abstract

Due to their ability to preferentially induce cell death in tumor cells, while sparing healthy cells, TNF-related apoptosis-inducing ligand (TRAIL) and agonistic anti-TRAIL-R1 or anti-TRAIL-R2-specific antibodies are under clinical investigations for cancer-treatment. However, TRAIL-Rs may also induce signaling pathways, which result in malignant progression. TRAIL receptors are transcriptionally upregulated via wild-type p53 following radio- or chemotherapy. Nevertheless, the impact of p53 status on the expression and signaling of TRAIL-Rs is not fully understood. Therefore, we analyzed side by side apoptotic and non-apoptotic signaling induced by TRAIL or the agonistic TRAIL-R-specific antibodies Mapatumumab (anti-TRAIL-R1) and Lexatumumab (anti-TRAIL-R2) in the two isogenic colon carcinoma cell lines HCT116 p53+/+ and p53-/-. We found that HCT116 p53+/+ cells were significantly more sensitive to TRAIL-R-triggering than p53-/- cells. Similarly, A549 lung cancer cells expressing wild-type p53 were more sensitive to TRAIL-R-mediated cell death than their derivatives with knockdown of p53. Our data demonstrate that the contribution of p53 in regulating TRAIL-R-induced apoptosis does not correlate to the levels of TRAIL-Rs at the plasma membrane, but rather to p53-mediated upregulation of Bax, favouring the mitochondrial amplification loop. Consistently, stronger caspase-9 and caspase-3 activation as well as PARP-cleavage was observed following TRAIL-R-triggering in HCT116 p53+/+ compared to HCT116 p53-/- cells. Interestingly, HCT116 p53+/+ cells showed also a more potent activation of non-canonical TRAIL-R-induced signal transduction pathways like JNK, p38 and ERK1/ERK2 than p53-/- cells. Likewise, these cells induced IL-8 expression in response to TRAIL, Mapatumumab or Lexatumumab significantly stronger than p53-/- cells. We obtained similar results in A549 cells with or without p53-knockdown and in the two isogenic colon cancer cell lines RKO p53+/+ and p53-/-. In both cellular systems, we could clearly demonstrate the potentiating effects of p53 on TRAIL-R-mediated IL-8 induction. In conclusion, we found that wild-type p53 increases TRAIL-R-mediated apoptosis but simultaneously augments non-apoptotic signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Apoptosis / physiology*
  • Cell Membrane / metabolism
  • Gene Knockdown Techniques
  • Genes, p53
  • HCT116 Cells
  • Humans
  • Interleukin-8 / biosynthesis
  • Neoplasms / metabolism*
  • Neoplasms / pathology
  • Receptor Activator of Nuclear Factor-kappa B / metabolism
  • Receptors, TNF-Related Apoptosis-Inducing Ligand / metabolism
  • Signal Transduction
  • TNF-Related Apoptosis-Inducing Ligand / metabolism*
  • Tumor Suppressor Protein p53 / deficiency
  • Tumor Suppressor Protein p53 / metabolism*
  • bcl-2-Associated X Protein / metabolism

Substances

  • BAX protein, human
  • CXCL8 protein, human
  • Interleukin-8
  • Receptor Activator of Nuclear Factor-kappa B
  • Receptors, TNF-Related Apoptosis-Inducing Ligand
  • TNF-Related Apoptosis-Inducing Ligand
  • TNFRSF10B protein, human
  • TNFRSF11A protein, human
  • TNFSF10 protein, human
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • bcl-2-Associated X Protein

Grants and funding

This work was supported by Deutsche Krebshilfe - Project 110788: Doktoranden-Stipendium des Mildred-Scheel-Doktorandenprogrammes given to Hella Schittek and Projekt 70112281 given to Anna Trauzold. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.