Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)

Casticin was obtained from natural plants, and it has been shown to exert biological functions; however, no report concerns the induction of DNA damage and repair in human lung cancer cells. The objective of this study was to investigate the effects and molecular mechanism of casticin on DNA damage...

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Main Authors: Zheng-Yu Cheng, Yung-Ting Hsiao, Yi-Ping Huang, Shu-Fen Peng, Wen-Wen Huang, Kuo-Ching Liu, Te-Chun Hsia, Tzong-Der Way, Jing-Gung Chung
Format: Article
Language:English
Published: MDPI AG 2020-01-01
Series:Molecules
Subjects:
Online Access:https://www.mdpi.com/1420-3049/25/2/341
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author Zheng-Yu Cheng
Yung-Ting Hsiao
Yi-Ping Huang
Shu-Fen Peng
Wen-Wen Huang
Kuo-Ching Liu
Te-Chun Hsia
Tzong-Der Way
Jing-Gung Chung
author_facet Zheng-Yu Cheng
Yung-Ting Hsiao
Yi-Ping Huang
Shu-Fen Peng
Wen-Wen Huang
Kuo-Ching Liu
Te-Chun Hsia
Tzong-Der Way
Jing-Gung Chung
author_sort Zheng-Yu Cheng
collection DOAJ
description Casticin was obtained from natural plants, and it has been shown to exert biological functions; however, no report concerns the induction of DNA damage and repair in human lung cancer cells. The objective of this study was to investigate the effects and molecular mechanism of casticin on DNA damage and repair in human lung cancer A549 cells. Cell viability was determined by flow cytometric assay. The DNA damage was evaluated by 4’,6-diamidino-2-phenylindole (DAPI) staining and electrophoresis which included comet assay and DNA gel electrophoresis. The protein levels associated with DNA damage and repair were analyzed by western blotting. The expression and translocation of p-H2A.X were observed by confocal laser microscopy. Casticin reduced total viable cell number and induced DNA condensation, fragmentation, and damage in A549 cells. Furthermore, casticin increased p-ATM at 6 h and increased p-ATR and BRCA1 at 6−24 h treatment but decreased p-ATM at 24−48 h, as well as decreased p-ATR and BRCA1 at 48 h. Furthermore, casticin decreased p-p53 at 6−24 h but increased at 48 h. Casticin increased p-H2A.X and MDC1 at 6−48 h treatment. In addition, casticin increased PARP (cleavage) at 6, 24, and 48 h treatment, DNA-PKcs and MGMT at 48 h in A549 cells. Casticin induced the expressions and nuclear translocation of p-H2AX in A549 cells by confocal laser microscopy. Casticin reduced cell number through DNA damage and condensation in human lung cancer A549 cells.
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spelling doaj.art-e3f64e47a6674c7e8f9f4270a9b7ea532022-12-22T01:28:12ZengMDPI AGMolecules1420-30492020-01-0125234110.3390/molecules25020341molecules25020341Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)Zheng-Yu Cheng0Yung-Ting Hsiao1Yi-Ping Huang2Shu-Fen Peng3Wen-Wen Huang4Kuo-Ching Liu5Te-Chun Hsia6Tzong-Der Way7Jing-Gung Chung8Department of Biological Science and Technology, China Medical University, Taichung 404, TaiwanDepartment of Biological Science and Technology, China Medical University, Taichung 404, TaiwanDepartment of Physiology, College of Medicine, China Medical University, Taichung 404, TaiwanDepartment of Biological Science and Technology, China Medical University, Taichung 404, TaiwanDepartment of Biological Science and Technology, China Medical University, Taichung 404, TaiwanDepartment of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 404, TaiwanDepartment of Respiratory Therapy, China Medical University, Taichung 404, TaiwanDepartment of Biological Science and Technology, China Medical University, Taichung 404, TaiwanDepartment of Biological Science and Technology, China Medical University, Taichung 404, TaiwanCasticin was obtained from natural plants, and it has been shown to exert biological functions; however, no report concerns the induction of DNA damage and repair in human lung cancer cells. The objective of this study was to investigate the effects and molecular mechanism of casticin on DNA damage and repair in human lung cancer A549 cells. Cell viability was determined by flow cytometric assay. The DNA damage was evaluated by 4’,6-diamidino-2-phenylindole (DAPI) staining and electrophoresis which included comet assay and DNA gel electrophoresis. The protein levels associated with DNA damage and repair were analyzed by western blotting. The expression and translocation of p-H2A.X were observed by confocal laser microscopy. Casticin reduced total viable cell number and induced DNA condensation, fragmentation, and damage in A549 cells. Furthermore, casticin increased p-ATM at 6 h and increased p-ATR and BRCA1 at 6−24 h treatment but decreased p-ATM at 24−48 h, as well as decreased p-ATR and BRCA1 at 48 h. Furthermore, casticin decreased p-p53 at 6−24 h but increased at 48 h. Casticin increased p-H2A.X and MDC1 at 6−48 h treatment. In addition, casticin increased PARP (cleavage) at 6, 24, and 48 h treatment, DNA-PKcs and MGMT at 48 h in A549 cells. Casticin induced the expressions and nuclear translocation of p-H2AX in A549 cells by confocal laser microscopy. Casticin reduced cell number through DNA damage and condensation in human lung cancer A549 cells.https://www.mdpi.com/1420-3049/25/2/341casticinhuman lung cancer a549 cellsdna damagedna condensation and repair
spellingShingle Zheng-Yu Cheng
Yung-Ting Hsiao
Yi-Ping Huang
Shu-Fen Peng
Wen-Wen Huang
Kuo-Ching Liu
Te-Chun Hsia
Tzong-Der Way
Jing-Gung Chung
Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)
Molecules
casticin
human lung cancer a549 cells
dna damage
dna condensation and repair
title Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)
title_full Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)
title_fullStr Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)
title_full_unstemmed Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)
title_short Casticin Induces DNA Damage and Affects DNA Repair Associated Protein Expression in Human Lung Cancer A549 Cells (Running Title: Casticin Induces DNA Damage in Lung Cancer Cells)
title_sort casticin induces dna damage and affects dna repair associated protein expression in human lung cancer a549 cells running title casticin induces dna damage in lung cancer cells
topic casticin
human lung cancer a549 cells
dna damage
dna condensation and repair
url https://www.mdpi.com/1420-3049/25/2/341
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