Premium
Knockout of VDAC1 in H9c2 Cells Promotes tBHP‐induced Cell Apoptosis Through Decreased Mitochondrial HK II Binding and Enhanced Glycolytic Stress
Author(s) -
Yang Meiying,
Li Keguo,
Sun Jie,
Stowe David F.,
Tajkhorshid Emad,
Kwok Wai-Meng,
Camara Amadou K.S.
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.06035
Subject(s) - vdac1 , microbiology and biotechnology , programmed cell death , mitochondrial permeability transition pore , mitochondrion , apoptosis , voltage dependent anion channel , biology , chemistry , cell growth , biochemistry , bacterial outer membrane , escherichia coli , gene
VDAC1, the most abundant mitochondrial outer membrane protein, transports ions and metabolites between the cytoplasm and mitochondria. It is also involved in cellular energy production and apoptosis, but the role of VDAC1 in cell death remains controversial. Silencing or upregulation of VDAC1 may stimulate or inhibit cell apoptosis, growth and survival depending on cell types and stimuli. Using VDAC1 knockout (Vdac1 −/− ) embryonic stem cells (ES) and mouse embryonic fibroblast (MEF), it was revealed that VDAC1 was dispensable for both mitochondrial membrane permeability transition pore (mPTP) opening and Bax and Bid‐driven cell death. The effect of Vdac1 −/− on oxidative stress‐induced cell death in a cardiac cell line has not been reported. We hypothesized that although VDAC1 is dispensable for mPTP and is not involved in mPTP‐mediated cell death, VDAC1 is an essential regulator of oxidative stress‐induced cell apoptosis in cardiac cells. To this end, we knocked out VDAC1 in the cardiac fibroblast cell line, H9c2 (Vdac1 −/− H9c2) by the CRISPR‐cas9 technique and examined whether VDAC1 plays a role in cell death via oxidative stress induced by tert‐butylhydroperoxide (tBHP), an organic peroxide. We found that under physiologic conditions, Vdac1 −/− did not affect H9c2 cell proliferation, and mitochondrial respiration. However, tBHP treatment enhanced the extracellular acidification rate (ECAR) and proton production rate (PPR) produced from glycolysis and promoted cell apoptosis in Vdac1 −/− H9c2 cells compared to the wildtype. Vdac1 −/− H9c2 cells also showed markedly reduced binding of mitochondria hexokinase II (HK II), a key glycolytic enzyme. Rescue of VDAC1 in Vdac1 −/− H9c2 cells reinstated mitochondria HK II binding and concomitantly decreased tBHP‐induced cell death. Mitochondrial respiration remained the same after tBHP treatment in Vdac1 −/− and wild type H9c2 cells. Our results suggest that Vdac1 −/− in H9c2 cells enhances tBHP‐induced oxidative stress‐mediated cell apoptosis by reducing mitochondria HK II binding and by increasing ECAR and PPR produced from glycolytic stress. Support or Funding Information National Institutes of Health (R01 HL‐131673‐01A1), Veterans Administration (BX‐002539‐01), United States.