β2-microglobulin induces caspase-dependent apoptosis in the CCRF-HSB-2 human leukemia cell line independently of the caspase-3, -8 and -9 pathways but through increased reactive oxygen species

John Gordon, Ching Huang Wu, Mojgan Rastegar, Ahmad R. Safa

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

Exogenous β2-microglobulin (β2m) induces significant apoptosis in the CCRF-HSB-2 human lymphoblastic leukemia cell line as detected by DNA fragmentation, DAP1 staining and annexin V binding assay. β2m treatment induced the release of cytochrome c and apoptosis-inducing factor (AIF) from the mitochondria, but no change in mitochondrial membrane potential (ΣΨm) was observed during apoptosis, suggesting that cytochrome c may be released through a mechanism independent of mitochondrial permeability transition (MPT) pore formation. Moreover, the β2m-induced release of cytochrome c and AIF from the mitochondria in CCRF-HSB-2 cells was caspase-independent, since Z-VAD-fmk, a general inhibitor of caspases, did not block the release of these factors. However, Z-VAD-fmk treatment significantly blocked β2m-induced apoptosis, while Western blot analysis revealed that caspases-1, -2, -3, -6, -7, -8 and -9 are not activated during β2m-induced apoptosis in these cells. These results collectively indicate that a post-mitochondrial caspase-dependent mechanism is involved in β2m-induced apoptosis. Moreover, β2m significantly enhanced the production of reactive oxygen species (ROS) during 12-48 hr treatment, and β2m-induced apoptosis was almost totally inhibited in cells pre-treated with the antioxidant N-acetylcysteine (NAC), providing evidence that β2m-induced apoptosis in CCRF-HSB-2 cells is ROS-dependent. Therefore, these results reveal that β2m-induced apoptosis in CCRF-HSB-2 cells may occur through an unknown caspase-dependent and ROS-dependent mechanism(s) that is associated with cytochrome c and AIF release from mitochondria, but is independent of the caspase -3, -8 and -9 pathways.

Original languageEnglish (US)
Pages (from-to)316-327
Number of pages12
JournalInternational Journal of Cancer
Volume103
Issue number3
DOIs
StatePublished - Jan 20 2003

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Keywords

  • β-microglobulin
  • Apoptosis
  • Caspases
  • cytochrome c
  • Leukemia
  • Mitochondria

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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