Empagliflozin drives ferroptosis in anoikis-resistant cells by activating miR-128-3p dependent pathway and inhibiting CD98hc in breast cancer

Free Radic Biol Med. 2024 Aug 1:220:288-300. doi: 10.1016/j.freeradbiomed.2024.05.018. Epub 2024 May 9.

Abstract

A tumour suppressor miRNA, miR-128-3p, is widely involved in various biological processes and has been found to get downregulated in breast cancer patients. We previously published that ectopically expressed miR-128-3p suppressed migration, invasion, cell cycle arrest, and breast cancer stem cells. In the present study, we explored the role of Empagliflozin (EMPA) as a miR-128-3p functionality-mimicking drug in inducing ferroptosis by inhibiting CD98hc. Given that CD98hc is one of the proteins critical in triggering ferroptosis, we confirmed that miR-128-3p and EMPA inhibited SP1, leading to inhibition of CD98hc expression. Further, transfection with siCD98hc, miR-128-3p mimics, and inhibitors was performed to assess their involvement in the ferroptosis of anoikis-resistant cells. We proved that anoikis-resistant cells possess high ROS and iron levels. Further, miR-128-3p and EMPA treatments induced ferroptosis by inhibiting GSH and enzymatic activity of GPX4 and also induced lipid peroxidation. Moreover, EMPA suppressed bioluminescence of 4T1-Red-FLuc induced thoracic cavity, peritoneal tumour burden and lung nodules in an in-vivo metastatic model of breast cancer. Collectively, we revealed that EMPA sensitized the ECM detached cells to ferroptosis by synergically activating miR-128-3p and lowering the levels of SP1 and CD98hc, making it a potential adjunct drug for breast cancer chemotherapy.

Keywords: 4T1 cells; Anoikis resistance; Breast cancer; Ferroptosis; Metastasis; Reactive oxygen species.

MeSH terms

  • Animals
  • Anoikis* / drug effects
  • Anoikis* / genetics
  • Benzhydryl Compounds* / pharmacology
  • Breast Neoplasms* / drug therapy
  • Breast Neoplasms* / genetics
  • Breast Neoplasms* / metabolism
  • Breast Neoplasms* / pathology
  • Cell Line, Tumor
  • Female
  • Ferroptosis* / drug effects
  • Ferroptosis* / genetics
  • Gene Expression Regulation, Neoplastic* / drug effects
  • Glucosides* / pharmacology
  • Humans
  • Lipid Peroxidation / drug effects
  • Mice
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Reactive Oxygen Species / metabolism
  • Sodium-Glucose Transporter 2 Inhibitors / pharmacology
  • Sodium-Phosphate Cotransporter Proteins, Type IIb
  • Xenograft Model Antitumor Assays

Substances

  • MicroRNAs
  • Glucosides
  • Benzhydryl Compounds
  • empagliflozin
  • MIRN128 microRNA, human
  • SLC34A2 protein, human
  • Reactive Oxygen Species
  • Sodium-Glucose Transporter 2 Inhibitors
  • Sodium-Phosphate Cotransporter Proteins, Type IIb