Reactive oxygen species generation and mitochondrial dysfunction in the apoptotic cell death of human myeloid leukemia HL-60 cells by a dietary compound withaferin A with concomitant protection by N-acetyl cysteine

Fayaz Malik, Ajay Kumar, Shashi Bhushan, Sheema Khan, Aruna Bhatia, Krishan Avtar Suri, Ghulam Nabi Qazi, Jaswant Singh

Research output: Contribution to journalArticle

162 Citations (Scopus)

Abstract

Induction of apoptosis in cancer cells has become the major focus of anti-cancer therapeutics development. WithaferinA, a major chemical constituent of Withania somnifera, reportedly shows cytotoxicity in a variety of tumor cell lines while its molecular mechanisms of action are not fully understood. We observed that withaferinA primarily induces oxidative stress in human leukemia HL-60 cells and in several other cancer cell lines. The withanolide induced early ROS generation and mitochondrial membrane potential (Δψ mt) loss, which preceded release of cytochrome c, translocation of Bax to mitochondria and apoptosis inducing factor to cell nuclei. These events paralleled activation of caspases -9, -3 and PARP cleavage. WA also activated extrinsic pathway significantly as evidenced by time dependent increase in caspase-8 activity vis-à-vis TNFR-1 over expression. WA mediated decreased expression of Bid may be an important event for cross talk between intrinsic and extrinsic signaling. Furthermore, withaferinA inhibited DNA binding of NF-κB and caused nuclear cleavage of p65/Rel by activated caspase-3. N-acetyl-cysteine rescued all these events suggesting thereby a pro-oxidant effect of withaferinA. The results of our studies demonstrate that withaferinA induced early ROS generation and mitochondrial dysfunction in cancer cells trigger events responsible for mitochondrial -dependent and -independent apoptosis pathways.

Original languageEnglish (US)
Pages (from-to)2115-2133
Number of pages19
JournalApoptosis
Volume12
Issue number11
DOIs
StatePublished - Nov 1 2007

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Acetylcysteine
Myeloid Leukemia
HL-60 Cells
Cell death
Cysteine
Reactive Oxygen Species
Cell Death
Cells
Caspase 3
Neoplasms
Withanolides
Withania
Apoptosis Inducing Factor
Apoptosis
Caspase 9
Caspase 8
Mitochondrial Membrane Potential
Cytochromes c
Tumor Cell Line
Cell Nucleus

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science
  • Clinical Biochemistry
  • Cell Biology
  • Biochemistry, medical
  • Cancer Research

Cite this

Reactive oxygen species generation and mitochondrial dysfunction in the apoptotic cell death of human myeloid leukemia HL-60 cells by a dietary compound withaferin A with concomitant protection by N-acetyl cysteine. / Malik, Fayaz; Kumar, Ajay; Bhushan, Shashi; Khan, Sheema; Bhatia, Aruna; Suri, Krishan Avtar; Qazi, Ghulam Nabi; Singh, Jaswant.

In: Apoptosis, Vol. 12, No. 11, 01.11.2007, p. 2115-2133.

Research output: Contribution to journalArticle

Malik, Fayaz ; Kumar, Ajay ; Bhushan, Shashi ; Khan, Sheema ; Bhatia, Aruna ; Suri, Krishan Avtar ; Qazi, Ghulam Nabi ; Singh, Jaswant. / Reactive oxygen species generation and mitochondrial dysfunction in the apoptotic cell death of human myeloid leukemia HL-60 cells by a dietary compound withaferin A with concomitant protection by N-acetyl cysteine. In: Apoptosis. 2007 ; Vol. 12, No. 11. pp. 2115-2133.
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