BAX-dependent mitochondrial pathway mediates the crosstalk between ferroptosis and apoptosis

Abstract

Ferroptosis is considered a distinctive form of cell death compared to other types of death such as apoptosis. It is known to result from iron-dependent accumulation of lipid peroxides rather than caspase activation. However, we reported recently that ferroptosis interplays with apoptosis. In this study, we investigated a possible mechanism of this interplay between ferroptosis and apoptosis. Results from our studies reveal that combined treatment of the ferroptotic agent erastin and the apoptotic agent TRAIL effectively disrupted mitochondrial membrane potential (ΔΨm) and subsequently promoted caspase activation. The alterations of mitochondrial membrane potential are probably due to an increase in oligomerization of BAX and its accumulation at the mitochondria during treatment with erastin and TRAIL. Interestingly, the combined treatment-promoted apoptosis was effectively inhibited in BAX-deficient HCT116 cells, but not BAK-deficient cells. These results indicate that the BAX-associated mitochondria-dependent pathway plays a pivotal role in erastin-enhanced TRAIL-induced apoptosis.

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Abbreviations

ATF4:

Activating transcription factor 4

BAK:

Bcl-2 homologous antagonist killer

BAX:

Bcl-2–associated X protein

Bcl-2:

B cell lymphoma 2

Bcl-xL:

B-cell lymphoma-extra large

BH3:

Bcl-2 homology

BID:

BH3 interacting-domain death agonist

C/EBP:

CCAAT-enhancer-binding proteins

CHOP:

CCAAT-enhancer-binding protein homologous protein

DAPI:

40,6-Diamidino-2-phenylindole

DISC:

Death-inducing signaling complex

DR:

Death receptor

DR4:

Death receptor 4

DR5:

Death receptor 5

EGTA:

Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

eIF2α:

Eukaryotic initiation factor 2α

ER:

Endoplasmic reticulum

ERA:

Erastin

GFP:

Green fluorescent protein

HB:

Homogenization buffer

HEPES:

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

HRP:

Horseradish peroxidase

JC-1:

5,50,6,60-Tetrachloro-1,10,3,30-tetraethylbenzimidazol carbocyanineiodid

PARP-1:

Poly [ADP-ribose] polymerase 1

PERK:

Protein kinase RNA-like endoplasmic reticulum kinase

PUMA:

p53 upregulated modulator of apoptosis

SD:

Standard deviation

TRAIL:

Tumor necrosis factor-related apoptosis-inducing ligand

WT:

Wild-type

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Acknowledgements

We thank Christine Burr (Department of Surgery, University of Pittsburgh) for her critical review of the manuscript.

Funding

Grant sponsor: NCI R03CA205267, R03CA212125, and P30CA047904

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Contributions

Y.S.L. K.K., and Y.S.P. were responsible for the data collection and analysis. X.L., M.H.A.C, D.L.B., and Y.J.L. were responsible for interpretation of the data. Y.S.L and Y.J.L. were responsible for the study concept and design. Y.S.L. and Y.J.L. were responsible for writing the manuscript.

Corresponding author

Correspondence to Yong J. Lee.

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The authors declare no competing financial interests.

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Lee, YS., Kalimuthu, K., Park, Y.S. et al. BAX-dependent mitochondrial pathway mediates the crosstalk between ferroptosis and apoptosis. Apoptosis 25, 625–631 (2020). https://doi.org/10.1007/s10495-020-01627-z

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Keywords

  • Ferroptosis
  • Apoptosis
  • Crosstalk
  • BAX
  • Mitochondria-dependent pathway