Role of Ca(2+) in diallyI disulfide-induced apoptotic cell death of HCT-15 cells..
- Author:
Eun Kyung PARK
1
;
Kang Beom KWON
;
Kwon In PARK
;
Byung Hyun PARK
;
Eun Chung JHEE
Author Information
1. Department of Biochemistry, Dental School, Chonbuk National University, Chonju, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
diallyl disulfide;
Ca2+;
H2O2;
caspase-3;
apoptosis;
HCT-15 cells
- MeSH:
Allyl Compounds/*pharmacology;
Apoptosis/*drug effects;
Calcium/*metabolism;
Caspases/metabolism;
Colonic Neoplasms/*metabolism/*pathology;
Disulfides/*pharmacology;
Enzyme Activation/drug effects;
Human;
Hydrogen Peroxide/metabolism;
Tumor Cells, Cultured
- From:Experimental & Molecular Medicine
2002;34(3):250-257
- CountryRepublic of Korea
- Language:English
-
Abstract:
Diallyl disulfide (DADS) induced apoptosis through the caspase-3 dependent pathway in leukemia cells was earlier reported from this laboratory. In this study, we investigated the involvement of Ca(2+) in DADS-induced apoptotic cell death of HCT-15, human colon cancer cell line. DADS induced the elevation of cytosolic Ca(2+) by biphasic pattern; rapid Ca(2+) peak at 3 min and following slow and sustained elevation till 3 h after the addition of DADS. Production of H(2)O(2) was also observed with its peak value at 4 h. Apoptotic pathways including the sequence of caspase-3 activation, poly(ADP-ribose) polymerase cleavage, and DNA fragmentation by DADS were completely blocked by various inhibitors such as specific caspase-3 inhibitor, free radical scavenger, and intracellular Ca(2+) chelator. N-acetylcystein and catalase treatment prevented the accumulation of H2O2 and later caspase-3 dependent apoptotic pathway. However, these radical scavengers did not block the elevation of intracellular Ca(2+). Treatment of cells with 1, 2-bis (2-aminophenoxyethane)-N, N, N-tetraacetic acid tetrakis -acetoxymethyl ester (BAPTA-AM), cellular Ca(2+) chelator, resulted in a complete blockage of the caspase-3 dependent apoptotic pathway of HCT-15 cells. It abolished the elevation of intracellular Ca(2+), and furthermore, completely inhibited the production of H(2)O(2). These results indicate that cytosolic Ca(2+) elevation is an earlier signaling event in apoptosis of HCT-15 cells. Collectively, our data demonstrate that DADS can induce apoptosis in HCT-15 cells through the sequential mechanism of Ca(2+) homeostasis disruption, accumulation of H(2)O(2), and resulting caspase-3 activation.
