Cantharidin induces G2/M phase arrest by inhibition of Cdc25c and Cyclin A and triggers apoptosis through reactive oxygen species and the mitochondria‑dependent pathways of A375.S2 human melanoma cells

Hsiao,Yu‑Ping; Tsai,Chung-Hung; Wu,Ping‑Ping; Hsu,Shu‑Chun; Liu,Hsin‑Chung; Huang,Yi‑Ping; Yang,Jen‑Hung; Chung,Jing‑Gung

doi:10.3892/ijo.2014.2689

Cantharidin induces G2/M phase arrest by inhibition of Cdc25c and Cyclin A and triggers apoptosis through reactive oxygen species and the mitochondria‑dependent pathways of A375.S2 human melanoma cells

Authors:
- Yu‑Ping Hsiao
- Chung-Hung Tsai
- Ping‑Ping Wu
- Shu‑Chun Hsu
- Hsin‑Chung Liu
- Yi‑Ping Huang
- Jen‑Hung Yang
- Jing‑Gung Chung
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Affiliations: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan, R.O.C., School of Pharmacy, China Medical University, Taichung, Taiwan, R.O.C., Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C., Department of Physiology, China Medical University, Taichung, Taiwan, R.O.C., Department of Dermatology, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, R.O.C.
Published online on: September 30, 2014 https://doi.org/10.3892/ijo.2014.2689
Pages: 2393-2402

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Abstract

Cantharidin (CTD), a component of natural mylabris (Mylabris phalerata Pallas) was reported to have high cytotoxicity in many human cancer cell lines. However, it was not reported to affect human melanoma A375.S2 cells. In the present study, we found that CTD induced cell morphological changes and decreased the percentage of viable cells and induced G2/M phase arrest and induction of apoptosis in A375.S2 cells. Results also showed that CTD induced the generation of reactive oxygen species (ROS) and Ca2+ and decreased mitochondria membrane potential and lead to the release of cytochrome c, AIF and Endo G. Further investigation revealed that CTD induced A375.S2 cells with an increase of caspase activation and caspase‑dependent apoptotic proteins to trigger correlated pathway mechanisms according to western blotting results. Western blotting was used for examining the changes of G2/M phase arrest and apoptosis‑associated protein expression and confocal laser microscopy was used to examine the translocation apoptosis‑associated protein. Results showed that CTD increased the protein expression of caspase‑3, ‑8 and ‑9, cytochrome c, Bax, Bid, Endo G and AIF but inhibited the levels of Bcl‑2 and Bcl‑x. CTD induced ER stress‑associated protein expression such as GRP78, IRE1β, ATF6α and caspase‑12. Based on those observations, we suggest that CTD may have potential as a novel anti‑cancer agent for the treatment of skin cancer.

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