Realgar-induced apoptosis of cervical cancer cell line Siha via cytochrome c release and caspase-3 and caspase-9 activation

Chin J Integr Med. 2012 May;18(5):359-65. doi: 10.1007/s11655-011-0697-z. Epub 2011 Apr 26.

Abstract

Objective: To explore the molecular mechanism of realgar-induced apoptosis of cervical cancer cells.

Methods: The cervical cancer cell line Siha was used to determine the cell viability and apoptosis after treatment with realgar using MTT assay and flow cytometry. The activities of caspase-3, -8, and -9 were detected by fluorescence resonance energy transfer technology and colorimetric assay, while the levels of Bcl-2, cytochrome c, and Bax were detected by Western blot method.

Results: Induction of apoptosis by realgar was detected in Siha cell line in a dose-dependent manner. The apoptosis was accompanied by a significant increase in cytochrome c release and activation of caspase-3 and caspase-9 but not caspase-8. Further, the realgar-induced apoptosis was inhibited by a broad-spectrum caspase inhibitor, a caspase-3 inhibitor, and a caspase-9 inhibitor but not by a caspase-8 inhibitor. Bcl-2 and Bax protein expressions were not changed by realgar.

Conclusion: The induction of apoptosis by realgar is mediated through a cytochrome c-dependent pathway, which sequentially activates caspase-9 and caspase-3.

MeSH terms

  • Apoptosis / drug effects*
  • Apoptosis / physiology
  • Arsenicals / pharmacology*
  • Caspase 3 / metabolism*
  • Caspase 8 / metabolism
  • Caspase 9 / metabolism*
  • Caspase Inhibitors
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Cytochromes c / metabolism*
  • Dose-Response Relationship, Drug
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Female
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Sulfides / pharmacology*
  • Uterine Cervical Neoplasms* / drug therapy
  • Uterine Cervical Neoplasms* / metabolism
  • Uterine Cervical Neoplasms* / pathology

Substances

  • Arsenicals
  • Caspase Inhibitors
  • Enzyme Inhibitors
  • Sulfides
  • arsenic disulfide
  • Cytochromes c
  • CASP3 protein, human
  • CASP8 protein, human
  • CASP9 protein, human
  • Caspase 3
  • Caspase 8
  • Caspase 9