miR-324-3p reverses cisplatin resistance by inducing GPX4-mediated ferroptosis in lung adenocarcinoma cell line A549

Biochem Biophys Res Commun. 2021 Apr 16;549:54-60. doi: 10.1016/j.bbrc.2021.02.077. Epub 2021 Mar 1.

Abstract

Purpose: MicroRNAs act as crucial regulators of a diverse range of biological processes, including chemoresistance. Our study aimed to investigate the effect of miR-324-3p on lung adenocarcinoma cell line A549 resistant to cis-diamminedichloroplatinum II (DDP, aka cisplatin).

Methods: The miR-324-3p expression levels in cisplatin-sensitive A549(A549) and cisplatin-resistant A549 (A549/DDP) cells were determined by qRT-PCR assay. Cell proliferation was determined with the commercial kit CCK-8 and colony formation assay, whereas cell death was analyzed using flow cytometry. The target gene of miR-324-3p was identified and validated with the luciferase reporter and western blot assays. The role of miR-324-3p in modulating cisplatin resistance was evaluated in vitro.

Results: The expression of miR-324-3p was found to be significantly downregulated in the A549/DDP cells. Conversely, miR-324-3p overexpression reversed cisplatin resistance in the cells. With regard to the possible mechanism underlying this phenomenon, we identified the glutathione peroxidase 4 (GPX4) gene as the direct target of miR-324-3p, where overexpression of the gene reversed the miR-324-3p effect of sensitizing the A549/DDP cells to cisplatin. Furthermore, the GPX4 inhibitor RSL3 could mimic the effect of miR-324-3p upregulation in increasing the sensitivity of the cisplatin-resistant cells to the drug. Significantly, miR-324-3p enhanced cisplatin-induced ferroptosis in the A549/DDP cells.

Conclusion: Our findings revealed the role of the miR-324-3p-GPX4 signaling axis in A549/DDP cells and how the targeting of this axis could be a potential strategy for reversing cisplatin resistance in human non small cell lung cancer (NSCLC).

Keywords: Cisplatin resistance; Ferroptosis; GPX4; NSCLC; miR-324-3p.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Adenocarcinoma of Lung / genetics*
  • Adenocarcinoma of Lung / pathology*
  • Adenocarcinoma of Lung / ultrastructure
  • Base Sequence
  • Cisplatin / pharmacology*
  • Down-Regulation / drug effects
  • Down-Regulation / genetics
  • Drug Resistance, Neoplasm / drug effects
  • Drug Resistance, Neoplasm / genetics*
  • Ferroptosis / drug effects
  • Ferroptosis / genetics*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / pathology
  • Lung Neoplasms / ultrastructure
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Mitochondria / drug effects
  • Mitochondria / ultrastructure
  • Phospholipid Hydroperoxide Glutathione Peroxidase / metabolism*
  • Up-Regulation / drug effects
  • Up-Regulation / genetics

Substances

  • MIRN324 microRNA, human
  • MicroRNAs
  • Phospholipid Hydroperoxide Glutathione Peroxidase
  • Cisplatin