ATP depletion alters the mode of cell death induced by benzyl isothiocyanate

Noriyuki Miyoshi, Etsuko Watanabe, Toshihiko Osawa, Masashi Okuhira, Yoshiyuki Murata, Hiroshi Ohshima, Yoshimasa Nakamura

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52 Citations (Scopus)


Pro-inflammatory death is presumably an undesirable event in cancer prevention process, thus biochemical comprehension and molecular definition of this process could have important clinical implications. In the present study, we examined the cytophysiological conversion of cell death mode by benzyl isothiocyanate (BITC) in human cervical cancer HeLa cells. The detailed studies using flow cytometric and morphological analyses demonstrated that the cells treated with appropriate concentration (25 μM) of BITC showed apoptotic feature, such as chromatin condensation, DNA fragmentation, and preserved plasma membrane integrity, whereas these features were disappeared by treatment with higher concentration (100 μM). The treatment with 2-deoxyglucose, an inhibitor of ATP synthesis, drastically increased in the ratio of necrotic dead cells, while it influences little that of apoptotic cells. Moreover, an analysis using the mitochondrial DNA-deficient HeLa cells demonstrated that the ρ° cells were more susceptible to the BITC-induced necrosis-like cell death compared to the wild-type (ρ+) cells, whereas the ROS production was significantly inhibited in the ρ° cells. It is likely that the BITC-induced ROS is derived from mitochondrial respiratory chain and ruled out the contribution to the mechanism of cell death mode switching. In addition, the BITC treatment resulted in a more rapid depletion of ATP in the ρ° cells than in the ρ+ cells. Furthermore, a caspase inhibitor, Z-VAD-fmk counteracted not only apoptosis, but also necrosis-like cell death induced by BITC, suggesting that increment in this cell death pattern might be due to the interruption of events downstream of a caspase-dependent pathway. The obtained data suggested that the decline in the intracellular ATP level plays an important role in tuning the mode of cell death by BITC.

Original languageEnglish
Pages (from-to)566-573
Number of pages8
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Issue number10
Publication statusPublished - Oct 2008


  • 2-DG
  • 2′,7′-dichlorofluorescin diacetate
  • AKR1C
  • ATM
  • ATP
  • Apoptosis
  • BITC
  • Benzyl isothiocyanate
  • CHX
  • ETP
  • GCLC
  • GPx
  • GSH
  • GST
  • HeLa cell
  • ITC
  • LDH
  • Necrosis
  • PI
  • ROS
  • TR
  • aldo-keto reductase 1C
  • ataxia telangiectasia-mutated
  • benzyl ITC
  • cycloheximide
  • etoposide
  • glutamate-cysteine ligase catalytic subunit
  • glutathione
  • glutathione peroxidase
  • glutathione-S-transferase
  • glyceraldehyde-3-phosphate dehydrogenase
  • isothiocyanate
  • lactate dehydrogenase
  • phenethyl ITC
  • propidium iodide
  • reactive oxygen species
  • thioredoxin

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology


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