Norcantharidin Induces Human Melanoma A375-S2 Cell Apoptosis through Mitochondrial and Caspase Pathways.
10.3346/jkms.2004.19.4.560
- Author:
Wei wei AN
1
;
Min wei WANG
;
Shin ichi TASHIRO
;
Satoshi ONODERA
;
Takashi IKEJIMA
Author Information
1. China-Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang, China. ikejimat@vip.sina.com
- Publication Type:Original Article
- Keywords:
Cantharidin;
Norcantharidin;
Cell Line;
Tumor;
A375-S2 Cells;
Apoptosis;
Caspase;
Mitochondria;
Proto-Oncogene Proteins c-bcl-2;
Cytochromes c
- MeSH:
Animals;
Apoptosis/*physiology;
Bicyclo Compounds, Heterocyclic/chemistry/metabolism/*pharmacology;
Caspases/antagonists & inhibitors/*metabolism;
Cell Line, Tumor/*drug effects;
Cell Shape;
DNA Fragmentation;
Enzyme Activation;
Humans;
Mitochondria/*metabolism;
Molecular Structure;
Proto-Oncogene Proteins c-bcl-2/metabolism;
Signal Transduction/*physiology
- From:Journal of Korean Medical Science
2004;19(4):560-566
- CountryRepublic of Korea
- Language:English
-
Abstract:
Norcantharidin (NCTD) is the demethylated form of cantharidin, which is the active substance of mylabris. To examine the pathway of NCTD-induced A375-S2 cell death, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-dipheyltetrazolium bromide (MTT) assay, photomicroscopical observation, DNA agarose gel electrophoresis, caspase activity assay and Western blot analysis were carried out. A375-S2 cells treated with NCTD exhibited several typical characteristics of apoptosis. The inhibitory effect of NCTD on human melanoma, A375-S2 cells, was partially reversed by the inhibitors of pan-caspase, caspase-3 and caspase-9. The activities of caspase-3 and -9 were significantly increased after treatment with NCTD at different time. The expression of inhibitor of caspase-activated DNase was decreased in a time-dependent manner, simultaneously, the ratio of Bcl-2/Bax or Bcl-xL/Bax was decreased and the expression ratio of proteins could be reversed by caspase-3 inhibitor. The expression of cytochrome c in cytosol was increased after NCTD treatment and caspase- 3 inhibitor had no significant effect on the up-regulation of cytochrom c. These results suggest that NCTD induced A375-S2 cell apoptosis and the activation of caspase and mitochondrial pathway were involved in the process of NCTD-induced A375-S2 cell apoptosis.
