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Int. J. Mol. Sci. 2017, 18(7), 1508; doi:10.3390/ijms18071508

Cudraflavone C Induces Apoptosis of A375.S2 Melanoma Cells through Mitochondrial ROS Production and MAPK Activation

1
Department of Nursing, Division of Basic Medical Sciences, Chang Gung University of Science and Technology, Chia-Yi 61363, Taiwan
2
Chronic Diseases and Health Promotion Research Center, Chang Gung University of Science and Technology, Chia-Yi 61363, Taiwan
3
Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, Taiwan
4
Department of Fragrance and Cosmetic Science, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
5
Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung 80424, Taiwan
6
Department of Pharmacy, College of Pharmacy & Health Care, Tajen University, Pingtung 90741, Taiwan
7
Center for Drug Abuse and Addiction, China Medical University Hospital, China Medical University, Taichung 40447, Taiwan
8
Division of Neurosurgery, Department of Surgery, Chang Gung Memorial Hospital, Chia-Yi61363, Taiwan
9
Department of Pediatrics, Division of Neonatology and Pediatric Hematology/Oncology, Chang Gung Memorial Hospital, Yunlin 63862, Taiwan
10
Department of Respiratory Care, Chang Gung University of Science and Technology, Chia-Yi Campus, Chia-Yi 61363, Taiwan
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 14 June 2017 / Revised: 4 July 2017 / Accepted: 6 July 2017 / Published: 13 July 2017
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

Melanoma is the most malignant form of skin cancer and is associated with a very poor prognosis. The aim of this study was to evaluate the apoptotic effects of cudraflavone C on A375.S2 melanoma cells and to determine the underlying mechanisms involved in apoptosis. Cell viability was determined using the MTT and real-time cytotoxicity assays. Flow cytometric evaluation of apoptosis was performed after staining the cells with Annexin V-FITC and propidium iodide. The mitochondrial membrane potential was evaluated using the JC-1 assay. Cellular ROS production was measured using the CellROX assay, while mitochondrial ROS production was evaluated using the MitoSOX assay. It was observed that cudraflavone C inhibited growth in A375.S2 melanoma cells, and promoted apoptosis via the mitochondrial pathway mediated by increased mitochondrial ROS production. In addition, cudraflavone C induced phosphorylation of MAPKs (p38, ERK, and JNK) and up-regulated the expression of apoptotic proteins (Puma, Bax, Bad, Bid, Apaf-1, cytochrome C, caspase-9, and caspase-3/7) in A375.S2 cells. Pretreatment of A375.S2 cells with MitoTEMPOL (a mitochondria-targeted antioxidant) attenuated the phosphorylation of MAPKs, expression of apoptotic proteins, and the overall progression of apoptosis. In summary, cudraflavone C induced apoptosis in A375.S2 melanoma cells by increasing mitochondrial ROS production; thus, activating p38, ERK, and JNK; and increasing the expression of apoptotic proteins. Therefore, cudraflavone C may be regarded as a potential form of treatment for malignant melanoma. View Full-Text
Keywords: cudraflavone C; mitochondria; melanoma cells; MAPKs; pro-oxidation; apoptosis cudraflavone C; mitochondria; melanoma cells; MAPKs; pro-oxidation; apoptosis
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MDPI and ACS Style

Lee, C.-W.; Yen, F.-L.; Ko, H.-H.; Li, S.-Y.; Chiang, Y.-C.; Lee, M.-H.; Tsai, M.-H.; Hsu, L.-F. Cudraflavone C Induces Apoptosis of A375.S2 Melanoma Cells through Mitochondrial ROS Production and MAPK Activation. Int. J. Mol. Sci. 2017, 18, 1508.

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