Casticin induces DNA damage and inhibits DNA repair-associated protein expression in B16F10 mouse melanoma cancer cells.

Abstract	Casticin, a polymethoxyflavone, has been demonstrated to possess anticancer activities, yet no study has shown in detail that casticin induces DNA damage in lung cancer cells. The purpose of this study was to investigate the possible molecular mechanisms of casticin which induce DNA damage and nuclear condensation in murine melanoma cancer B16F10 cells. In this study, by examining and capturing images using phase contrast microscopy, we found that casticin induced cell morphological changes. Moreover, it decreased the total number of viable cells which was measured by flow cytometry. Casticin-induced DNA damage and nuclear DNA condensation were measured by DAPI staining, respectively. Western blotting indicated that casticin decreased the protein levels of O6‑methylguanine-DNA methyltransferase (MGMT), breast cancer 1, early onset (BRCA1), mediator of DNA damage checkpoint 1 (MDC1), DNA-dependent protein kinase (DNA-PK) but increased phospho-p53 tumor suppressor protein (p-p53), phospho-ataxia telangiectasia mutated kinase (p-ATM), phospho-histone H2A.X (Ser139) and poly(ADP-ribose) polymerase (PARP) in the B16F10 cells. Furthermore, we used confocal laser system microscopy to examine the protein expression levels and we found that casticin increased the expression of p-p53 and p-H2A.X in the B16F10 cells. Collectively, casticin induced DNA damage and affected DNA repair proteins in the B16F10 cells in vitro.
Authors	Yung-Luen Shih, Jason Chou, Ming-Yang Yeh, Hsiao-Min Chou, Hsiu-Chen Chou, Hsu-Feng Lu, Hung-Sheng Shang, Fu-Shin Chueh, Yung-Lin Chu, Shu-Ching Hsueh, Jing-Gung Chung
Journal	Oncology reports (Oncol Rep) Vol. 36 Issue 4 Pg. 2094-100 (Oct 2016) ISSN: 1791-2431 [Electronic] Greece
PMID	27572101 (Publication Type: Journal Article)
Chemical References	Antineoplastic Agents, Phytogenic Flavonoids casticin
Topics	Animals Antineoplastic Agents, Phytogenic (pharmacology) Blotting, Western Cell Line, Tumor DNA Damage (drug effects) DNA Repair (drug effects) Disease Models, Animal Flavonoids (pharmacology) Flow Cytometry Melanoma Mice Microscopy, Confocal Microscopy, Phase-Contrast

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