Reactive oxygen species-dependent apoptosis induction by water extract of Citrus unshiu peel in MDA-MB-231 human breast carcinoma cells
2
조회
0
다운로드
4
인용
0
참고문헌
키워드
초록
BACKGROUND/OBJECTIVES: Although several recent studies have reported the anti-cancer effects of extracts or components of Citrus unshiu peel, which has been used for various purposes in traditional medicine, the molecular mechanisms for their effects remain unclear. In the present study, the anti-cancer activity of a water-soluble extract of C. unshiu peel (WECU) in MDA-MB-231 human breast carcinoma cells at the level of apoptosis induction was investigated. MATERIALS/METHODS: Cytotoxicity was evaluated using the MTT assay. Apoptosis was detected using DAPI staining and flow cytometry analyses. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, caspase activity and Western blotting were used to confirm the basis of apoptosis. RESULTS: The results indicated that WECU-induced apoptosis was related to the activation of caspase-8, and -9, representative initiator caspases of extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3 accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and down-regulation of the inhibitors of apoptosis protein family members. WECU also increased the pro-apoptotic BAX to anti-apoptotic BCL-2 ratio, loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm. Furthermore, WECU provoked the generation of ROS, but the reduction of cell viability and induction of apoptosis by WECU were prevented when ROS production was blocked by antioxidant N-acetyl cysteine. CONCLUSIONS: These results suggest that WECU suppressed proliferation of MDA-MB-231 cells by activating extrinsic and intrinsic apoptosis pathways in a ROS-dependent manner.
Abstract
BACKGROUND/OBJECTIVES: Although several recent studies have reported the anti-cancer effects of extracts or components of Citrus unshiu peel, which has been used for various purposes in traditional medicine, the molecular mechanisms for their effects remain unclear. In the present study, the anti-cancer activity of a water-soluble extract of C. unshiu peel (WECU) in MDA-MB-231 human breast carcinoma cells at the level of apoptosis induction was investigated. MATERIALS/METHODS: Cytotoxicity was evaluated using the MTT assay. Apoptosis was detected using DAPI staining and flow cytometry analyses. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, caspase activity and Western blotting were used to confirm the basis of apoptosis. RESULTS: The results indicated that WECU-induced apoptosis was related to the activation of caspase-8, and -9, representative initiator caspases of extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3 accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and down-regulation of the inhibitors of apoptosis protein family members. WECU also increased the pro-apoptotic BAX to anti-apoptotic BCL-2 ratio, loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm. Furthermore, WECU provoked the generation of ROS, but the reduction of cell viability and induction of apoptosis by WECU were prevented when ROS production was blocked by antioxidant N-acetyl cysteine. CONCLUSIONS: These results suggest that WECU suppressed proliferation of MDA-MB-231 cells by activating extrinsic and intrinsic apoptosis pathways in a ROS-dependent manner.
참고문헌 (27)
- Siegel RL, "Cancer statistics, 2016", CA Cancer J Clin, 66, pp.7~30 (2016)[DOI]
- Niraula S, "Mechanism of drug resistance in relation to site of metastasis: meta-analyses of randomized controlled trials in advanced breast cancer according to anticancer strategy", Cancer Treat Rev, 50, pp.168~174 (2016)[DOI]
- Li C, "Systematic review and meta-analysis suggest that dietary cholesterol intake increases risk of breast cancer", Nutr Res, 36, pp.627~635 (2016)[DOI]
- Safarzadeh E, "Herbal medicine as inducers of apoptosis in cancer treatment", Adv Pharm Bull, 4, pp.421~427 (2014)
- Gali-Muhtasib H, "Cell death mechanisms of plant-derived anticancer drugs: beyond apoptosis", Apoptosis, 20, pp.1531~1562 (2015)[DOI]
- Bonofiglio D, "Natural products as promising antitumoral agents in breast cancer: mechanisms of action and molecular targets", Mini Rev Med Chem, 16, pp.596~604 (2016)[DOI]
- Wang CY, "Traditional Chinese medicine: a treasured natural resource of anticancer drug research and development", Am J Chin Med, 42, pp.543~559 (2014)
- Ming-Hua C, "Mechanisms of anorexia cancer cachexia syndrome and potential benefits of traditional medicine and natural herbs", Curr Pharm Biotechnol, 17, pp.1147~1152 (2016)[DOI]
- Min KY, "Antimicrobial activity of acid-hydrolyzed Citrus unshiu peel extract in milk", J Dairy Sci, 97, pp.1955~1960 (2014)[DOI]
- Park HJ, "Citrus unshiu peel extract ameliorates hyperglycemia and hepatic steatosis by altering inflammation and hepatic glucose- and lipid-regulating enzymes in db", db mice, 2013, pp.24, 419~427
- Oh YC, "Anti-inflammatory effect of Citrus unshiu peel in LPS-stimulated RAW 264.7 macrophage cells", Am J Chin Med, 40, pp.611~629 (2012)[DOI]
- Suzuki M, "Anti-hepatitis C virus effect of Citrus unshiu peel and its active ingredient nobiletin", Am J Chin Med, 33, pp.87~94 (2005)[DOI]
- Lee S, "Extracts from Citrus unshiu promote immune-mediated inhibition of tumor growth in a murine renal cell carcinoma model", J Ethnopharmacol, 133, pp.973~979 (2011)[DOI]
- Kim A, "Citrus unshiu peel extract alleviates cancer-induced weight loss in mice bearing CT-26 adenocarcinoma", Sci Rep, 6, pp.24214~ (2016)[DOI]
- Jin H, "Flavonoids from Citrus unshiu Marc. inhibit cancer cell adhesion to endothelial cells by selective inhibition of VCAM-1", Oncol Rep, 30, pp.2336~2342 (2013)[DOI]
- Park HR, "Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu)", Int J Biol Macromol, 94, pp.161~169 (2017)
- 이필영, "Histone H4 is cleaved by granzyme A during staurosporine-induced cell death in B-lymphoid Raji cells", BMB Reports, 49(10), pp.560~565 (2016), 생화학분자생물학회
- 유미경, "Pear pomace ethanol extract improves insulin resistance through enhancement of insulin signaling pathway without lipid accumulation", Nutrition Research and Practice, 11(3), pp.198~205 (2017), 한국영양학회
- 김한빛, "Propolis Inhibits UVA-Induced Apoptosis of Human Keratinocyte HaCaT Cells by Scavenging ROS", 한국독성학회지, 32(4), pp.345~351 (2016), 한국독성학회
- Fulda S, "Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy", Oncogene, 25, pp.4798~4811 (2006)[DOI]
- Nakajima YI, "Caspase-dependent non-apoptotic processes in development", Cell Death Differ, 24, pp.1422~1430 (2017)[DOI]
- Decker P, "Modulating poly (ADP-ribose) polymerase activity: potential for the prevention and therapy of pathogenic situations involving DNA damage and oxidative stress", Curr Pharm Biotechnol, 3, pp.275~283 (2002)
- Hata AN, "The BCL2 family: Key mediators of the apoptotic response to targeted anticancer therapeutics", Cancer Discov, 5, pp.475~487 (2015)
- Kaminskyy VO, "Free radicals in cross talk between autophagy and apoptosis", Antioxid Redox Signal, 21, pp.86~102 (2014)[DOI]
- Yang Y, "Mitochondria and mitochondrial ROS in cancer: novel targets for anticancer therapy", J Cell Physiol, 231, pp.2570~2581 (2016)[DOI]
- 김종두, "ROS homeostasis and metabolism: a critical liaison for cancer therapy", Experimental and Molecular Medicine, 48, pp.1~13 (2016), 생화학분자생물학회
- Kim B, "Mitochondrial dynamics altered by oxidative stress in cancer", Free Radic Res, 50, pp.1065~1070 (2016)[DOI]