The PTBP1‑NCOA4 axis promotes ferroptosis in liver cancer cells

Oncol Rep. 2023 Feb;49(2):45. doi: 10.3892/or.2023.8482. Epub 2023 Jan 20.

Abstract

Polypyrimidine tract‑binding protein 1 (PTBP1) plays an important role in tumor immunity, cell proliferation, apoptosis, and autophagy by regulating RNA metabolism. However, the specific function and mechanism of PTBP1 in ferroptosis remain unclear. In the present study, it was investigated whether PTBP1 regulates ferroptosis and the exact mechanism. The iron, malondialdehyde (MDA), and GSH levels were detected in sorafenib (SF)‑treated liver cancer cells. si‑PTBP1 introduction into SF‑treated liver cancer cells resulted in a significant reduction in the levels of MDA and iron. Additionally, a significant recovery of GSH levels was observed after silencing PTBP1. StarBase v2.0 database was used to predict potential transcripts that can physically interact with PTBP1 and nuclear receptor coactivator 4 (NCOA4) mRNA was identified as the most enriched binding partner in the PTBP1‑RNA complex. A dual‑luciferase assay then demonstrated that PTBP1 directly interacted with NCOA4. PTBP1 silencing did not affect NCOA4 stability following treatment with cycloheximide. A pull‑down assay revealed that the PTBP1‑binding region was in the 5'‑UTR of the NCOA4 mRNA sequence. These results suggest that PTBP1 mediates ferroptosis in liver cancer cells by regulating NCOA4 translation. In vivo experiments reconfirmed the role of the PTBP1‑NCOA4 axis in a xenograft transplantation model. It was observed that the mean tumor weight increased after PTBP1 knockout. In conclusion, silencing of PTBP1 decreased the sensitivity of liver cancer cells to ferroptosis after SF treatment and regulated ferritinophagy by mediating NCOA4 translation.

Keywords: NCOA4; PTBP1; RNA‑binding protein; ferritinophagy; ferroptosis.

MeSH terms

  • Animals
  • Autophagy / genetics
  • Ferroptosis* / genetics
  • Heterogeneous-Nuclear Ribonucleoproteins / genetics
  • Heterogeneous-Nuclear Ribonucleoproteins / metabolism
  • Humans
  • Iron / metabolism
  • Liver Neoplasms* / genetics
  • Liver Neoplasms* / pathology
  • Nuclear Receptor Coactivators / metabolism
  • Polypyrimidine Tract-Binding Protein / genetics
  • Polypyrimidine Tract-Binding Protein / metabolism
  • RNA
  • RNA, Messenger / genetics
  • Sorafenib
  • Transcription Factors / metabolism

Substances

  • Heterogeneous-Nuclear Ribonucleoproteins
  • Iron
  • NCOA4 protein, human
  • Nuclear Receptor Coactivators
  • Polypyrimidine Tract-Binding Protein
  • PTBP1 protein, human
  • RNA
  • RNA, Messenger
  • Sorafenib
  • Transcription Factors

Grants and funding

The present study was supported by the National Natural Science Foundation of China (grant nos. 82172830 and 82002504), and the Shandong Provincial Key Research and Development Program (grant no. 2019GSF108053).