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

Shih,Yung-Luen; Chou,Jason; Yeh,Ming-Yang; Chou,Hsiao-Min; Chou,Hsiu-Chen; Lu,Hsu-Feng; Shang,Hung-Sheng; Chueh,Fu-Shin; Chu,Yung-Lin; Hsueh,Shu-Ching; Chung,Jing-Gung

doi:10.3892/or.2016.5027

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

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
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Affiliations: Department of Pathology and Laboratory Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan, R.O.C., Department of Pathology, Cheng-Hsin General Hospital, Taipei, Taiwan, R.O.C., Office of Director, Cheng-Hsin General Hospital, Taipei, Taiwan, R.O.C., Department of Restaurant, Hotel and Institutional Management, Fu-Jen Catholic University, New Taipei, Taiwan, R.O.C., Department of Pathology, National Defense Medical Center, Division of Clinical Pathology, Tri-Service General Hospital, Taipei, Taiwan, R.O.C., Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan, R.O.C., International Master's Degree Program in Food Science, International College, National Pingtung University of Science and Technology, Pingtung, Taiwan, R.O.C., Department of Clinical Pathology, Cheng-Hsin General Hospital, Taipei, Taiwan, R.O.C., Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
Published online on: August 17, 2016 https://doi.org/10.3892/or.2016.5027
Pages: 2094-2100

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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.

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