Exp Mol Med.  2003 Apr;35(2):83-90.

Arsenic trioxide-induced apoptosis is independent of stress-responsive signaling pathways but sensitive to inhibition of inducible nitric oxide synthase in HepG2 cells

• Kang SH
• Song JH
• Kang HK
• Kang JH
• Kim SJ
• Kang HW
• Lee YK
• Park DB

Abstract

Arsenic trioxide (As2O3) has been found to be remarkably effective in the treatment of patients with acute promyelocytic leukemia (APL). Although evidences for the proapoptotic activity of As2O3 have been suggested in leukemic and other solid cancer cells, the nature of intracellular mechanisms is far from clear. In the present study, we investigated As2O3 affect on the stress-responsive signaling pathways and pretreatment with antioxidants using HepG2 cells. When treated with micromolar concentrations of As2O3, HepG2 cells became highly apoptotic paralleled with activation of caspase-3 and members of mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase (ERK) and c-jun NH2-terminal kinase (JNK) but not p38 MAP kinase. However, inhibition of each kinase activity failed to inhibit apoptosis by As2O3. Addition of n-acetyl cysteine (NAC) or diphenyleneiodonium (DPI) effectively protected cells from apoptosis and significantly lowered As2O3-induced activation of caspase-3. However, neither NAC nor DPI was able to effect ERK or JNK activation induced by As2O3. Guanidinoethyldisulfide dihydrochloride (GED) and 2-ethyl- 2-thiopseudourea (ETU), known inhibitors of the inducible nitric oxide synthase (iNOS), also suppressed the apoptotic activity of As2O3. These results suggest that As2O3 induces caspase-mediated apoptosis involving a mechanism generating oxidative stress. However, activation of some stress- responsive signaling pathways by As2O3 may not be the major determinant in the course of apoptotic processes.

Keyword

Keywords; antioxidant; apoptosis; arsenicals; extra-cellular signal-regulated kinase; mitogen-activated protein kinase; nitric oxide synthase

MeSH Terms

Antioxidants/administration & dosage/pharmacology
Apoptosis/*drug effects
Arsenicals/administration & dosage/*pharmacology
Cell Line, Tumor
Dose-Response Relationship, Drug
Enzyme Activation/drug effects
Enzyme Inhibitors/*pharmacology
Human
Nitric-Oxide Synthase/*antagonists & inhibitors/metabolism
*Oxidative Stress/drug effects
Oxides/administration & dosage/*pharmacology
*Signal Transduction/drug effects