Experimental and Molecular Medicine

Korean Society for Biochemistry and Molecular Biongy (생화학분자생물학회)
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Role of $Ca^{2+}$ in diallyl disulfide-induced apoptotic cell death of HCT-15 cells

Park, Eun-Kyung;Kwon, Kang-Beom;Park, Kie-In;Jhee, Eun-Chung

  • Published : 2002.07.31
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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 $H_2O_2$ 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 $H_2O_2$ 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 $H_2O_2$. 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 $H_2O_2$, and resulting caspase-3 activation.

Keywords

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