Ferroptosis has been previously implicated in the pathological progression of
cardiomyopathy.
Herceptin (
trastuzumab), which targets HER2, is commonly applied for the treatment of HER2+
breast cancer. However, its clinical use is limited by its
cardiotoxicity. Therefore, the present study aimed to investigate if targeting ferroptosis could protect against Herceptin‑induced
heart failure in an in vitro model of H9c2 cells
after treatment of
Herceptin, Herceptin + ferroptosis inhibitor ferrostatin‑1 (Fer‑1) or Herceptin +
Deferoxamine. H9c2 cell viability was measured by MTT assay.
Reactive oxygen species (ROS) levels were detected by measuring the fluorescence of DCFH‑DA‑A and MitoSOX™ Red.
Glutathione (GSH)/
oxidized glutathione (
GSSG) ratio was measured using the GSH/
GSSG Ratio Detection Assay kit. Mitochondrial membrane potential and
ATP content were evaluated by JC‑1 staining and bioluminescent assay kits, respectively.
Protein expressions of
glutathione peroxidase 4, recombinant solute carrier family 7 member 11, mitochondrial optic atrophy1‑1/2, mitofusin, Acyl‑CoA
synthetase long chain family member 4, cytochrome c, voltage‑dependent anion‑selective channel, dynamin‑related
protein, mitochondrial fission 1
protein and mitochondrial
ferritin were evaluated by western blotting. It was found that
Herceptin reduced H9c2 cell viability whilst increasing intracellular and mitochondrial ROS levels in a dose‑ and time‑dependent manner. Furthermore,
Herceptin decreased
glutathione peroxidase (GPX)
protein expression and the GSH/
GSSG ratio in H9c2 cells in a dose‑ and time‑dependent manner. The Fer‑1 abolished this Herceptin‑induced reduction in cell viability, GSH/
GSSG ratio, mitochondrial membrane potential and
ATP content. Fer‑1 also reversed the suppressive effects of
Herceptin on the
protein expression levels of GPX4, recombinant solute carrier family 7 member 11, mitochondrial optic atrophy1‑1/2 and mitofusin in H9c2 cells. Subsequently, Fer‑1 was found to reverse the Herceptin‑induced increase in mitochondrial ROS and
iron levels in H9c2 cells, as well as the increased
protein expression levels of Acyl‑CoA
synthetase long chain family member 4, cytochrome c, voltage‑dependent anion‑selective channel, dynamin‑related
protein, mitochondrial fission 1
protein and mitochondrial
ferritin in H9c2 cells. However, compared with
deferoxamine, an
iron chelator, the effects of Fer‑1 were less effective. Collectively, these findings provided insights into the pathogenic mechanism that underlie Herceptin‑induced
cardiomyopathy, which potentially provides a novel therapeutic target for the prevention of
cardiotoxicity in HER2+
breast cancer treatment.