Dong -KYU MOON B, Dong- b HEE KİM, Kwang -DONG KİM, Sun -CHUL HWANG, Young- JİN PARK B, Dong- YEONG LEE

Cytotoxic effects of delphinidin in human osteosarcoma cells

Introduction: The aim of this study was to evaluate whether delphinidin is cytoprotective or cytotoxic inosteosarcoma cell lines, and to elucidate the underlying mechanisms.Materials and methods: The present study investigated whether apoptosis or autophagy is induced bydelphinidin in human osteosarcoma cell lines. Delphinidin was used as the antioxidant, along with twoautophagy inhibitors: 3-methyladenine and baŞlomycin A1. Cell viability and known autophagicmarkers, such as LC3-II expression, were evaluated. Reactive oxygen species (ROS) formation and cellcycle analysis were also investigated.Results: Delphinidin showed concentration-dependent cytotoxicity to osteosarcoma cell. Delphinidin isclosely associated with apoptotic cell death mechanisms and pathways related to ROS accumulation. Inaddition, we observed delphinidin-induced autophagosome formation and increasing levels of LC3-IIconversion. However, in spite of delphinidin induced autophagy, the cytotoxic effects induced in theosteosarcoma cells may not be operating via autophagic cell death mechanisms.Conclusions: Delphinidin compromises the cellular protective mechanisms by inhibiting autophagy,permitting ROS to accumulate andŞnally enhance apoptotic cell death. Our results indicate that delphinidin may play a critical role as a chemotherapeutic agent by preventing the development andprogression of osteosarcoma cells.© 2017 Turkish Association of Orthopaedics and Traumatology. Publishing services by Elsevier B.V. This isan open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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1. Bacci G, Longhi A, Versari M, Mercuri M, Briccoli A, Picci P. Prognostic factors for osteosarcoma of the extremity treated with neoadjuvant chemotherapy: 15- year experience in 789 patients treated at a single institution. Cancer. 2006;106:1154e1161.

2. Kong JM, Chia LS, Goh NK, Chia TF, Brouillard R. Analysis and biological activities of anthocyanins. Phytochemistry. 2003;64:923e933.

3. Cvorovic J, Tramer F, Granzotto M, Candussio L, Decorti G, Passamonti S. Oxidative stress-based cytotoxicity of delphinidin and cyanidin in colon cancer cells. Arch Biochem Biophys. 2010;501:151e157.

4. Lim W, Jeong W, Song G. Delphinidin suppresses proliferation and migration of human ovarian clear cell carcinoma cells through blocking AKT and ERK1/2 MAPK signaling pathways. Mol Cell Endocrinol. 2016;422:172e181.

5. Yun JM, Afaq F, Khan N, Mukhtar H. Delphinidin, an anthocyanidin in pigmented fruits and vegetables, induces apoptosis and cell cycle arrest in human colon cancer HCT116 cells. Mol Carcinog. 2009;48:260e270.

6. Tsuyuki S, Fukui S, Watanabe A, Akune S, Tanabe M, Yoshida K. Delphinidin induces autolysosome as well as autophagosome formation and delphinidininduced autophagy exerts a cell protective role. J Biochem Mol Toxicol. 2012;26: 445e453.

7. Li J, Yang Z, Li Y, et al. Cell apoptosis, autophagy and necroptosis in osteosarcoma treatment. Oncotarget. 2016;7:44763e44778.

8. Yang ZJ, Chee CE, Huang S, Sinicrope FA. The role of autophagy in cancer: therapeutic implications. Mol Cancer Ther. 2011;10:1533e1541.

9. Park SH, Kim JH, Chi GY, et al. Induction of apoptosis and autophagy by sodium selenite in A549 human lung carcinoma cells through generation of reactive oxygen species. Toxicol Lett. 2012;212:252e261.

10. Cherukuri DP, Nelson MA. Role of reactive oxygen species (ROS) and JNKs in selenite-induced apoptosis in HepG2 cells. Cancer Biol Ther. 2008;7:697e698.

11. Scherz-Shouval R, Shvets E, Fass E, Shorer H, Gil L, Elazar Z. Reactive oxygen species are essential for autophagy and specifically regulate the activity of Atg4. EMBO J. 2007;26:1749e1760.

12. Feng R, Ni HM, Wang SY, et al. Cyanidin-3-rutinoside, a natural polyphenol antioxidant, selectively kills leukemic cells by induction of oxidative stress. J Biol Chem. 2007;282:13468e13476.

13. Duan Y, Ke J, Zhang H, He Y, Sun G, Sun X. Autophagic cell death of human hepatoma G2 cells mediated by procyanidins from Castanea mollissima Bl. Shell-induced reactive oxygen species generation. Chem Biol Interact. 2014;224:13e23.

14. Mathew R, Karp CM, Beaudoin B, et al. Autophagy suppresses tumorigenesis through elimination of p62. Cell. 2009;137:1062e1075.

15. Macip S, Igarashi M, Berggren P, Yu J, Lee SW, Aaronson SA. Influence of induced reactive oxygen species in p53-mediated cell fate decisions. Mol Cell Biol. 2003;23:8576e8585.

16. Galluzzi L, Maiuri MC, Vitale I, et al. Cell death modalities: classification and pathophysiological implications. Cell Death Differ. 2007;14:1237e1243.

17. Ling YH, Aracil M, Zou Y, et al. PM02734 (elisidepsin) induces caspase-independent cell death associated with features of autophagy, inhibition of the Akt/ mTOR signaling pathway, and activation of death-associated protein kinase. Clin Cancer Res. 2011;17:5353e5366.

18. Hafeez BB, Siddiqui IA, Asim M, et al. A dietary anthocyanidin delphinidin induces apoptosis of human prostate cancer PC3 cells in vitro and in vivo: involvement of nuclear factor-kappaB signaling. Cancer Res. 2008;68: 8564e8572.

19. Vermeulen K, Van Bockstaele DR, Berneman ZN. The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif. 2003;36: 131e149.

20. Bucher N, Britten CD. G2 checkpoint abrogation and checkpoint kinase-1 targeting in the treatment of cancer. Br J Cancer. 2008;98:523e528.

21. Meiers S, Kemeny M, Weyand U, Gastpar R, von Angerer E, Marko D. The anthocyanidins cyanidin and delphinidin are potent inhibitors of the epidermal growth-factor receptor. J Agric Food Chem. 2001;49:958e962.

22. Duluc L, Jacques C, Soleti R, Andriantsitohaina R, Simard G. Delphinidin inhibits VEGF induced-mitochondrial biogenesis and Akt activation in endothelial cells. Int J Biochem Cell Biol. 2014;53:9e14.