Withaferin A Induces Oxidative Stress-Mediated Apoptosis and DNA Damage in Oral Cancer Cells
Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We...
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| Published in | Frontiers in physiology Vol. 8; p. 634 |
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| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English |
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07.09.2017
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| Abstract | Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We evaluated whether the killing ability of WFA is selective, and we explored its mechanism against oral cancer cells. An MTS tetrazolium cell proliferation assay confirmed that WFA selectively killed two oral cancer cells (Ca9-22 and CAL 27) rather than normal oral cells (HGF-1). WFA also induced apoptosis of Ca9-22 cells, which was measured by flow cytometry for subG1 percentage, annexin V expression, and pan-caspase activity, as well as western blotting for caspases 1, 8, and 9 activations. Flow cytometry analysis shows that WFA-treated Ca9-22 oral cancer cells induced G2/M cell cycle arrest, ROS production, mitochondrial membrane depolarization, and phosphorylated histone H2A.X (γH2AX)-based DNA damage. Moreover, pretreating Ca9-22 cells with
-acetylcysteine (NAC) rescued WFA-induced selective killing, apoptosis, G2/M arrest, oxidative stress, and DNA damage. We conclude that WFA induced oxidative stress-mediated selective killing of oral cancer cells. |
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| AbstractList | Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We evaluated whether the killing ability of WFA is selective, and we explored its mechanism against oral cancer cells. An MTS tetrazolium cell proliferation assay confirmed that WFA selectively killed two oral cancer cells (Ca9-22 and CAL 27) rather than normal oral cells (HGF-1). WFA also induced apoptosis of Ca9-22 cells, which was measured by flow cytometry for subG1 percentage, annexin V expression, and pan-caspase activity, as well as western blotting for caspases 1, 8, and 9 activations. Flow cytometry analysis shows that WFA-treated Ca9-22 oral cancer cells induced G2/M cell cycle arrest, ROS production, mitochondrial membrane depolarization, and phosphorylated histone H2A.X (γH2AX)-based DNA damage. Moreover, pretreating Ca9-22 cells with
-acetylcysteine (NAC) rescued WFA-induced selective killing, apoptosis, G2/M arrest, oxidative stress, and DNA damage. We conclude that WFA induced oxidative stress-mediated selective killing of oral cancer cells. Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We evaluated whether the killing ability of WFA is selective, and we explored its mechanism against oral cancer cells. An MTS tetrazolium cell proliferation assay confirmed that WFA selectively killed two oral cancer cells (Ca9-22 and CAL 27) rather than normal oral cells (HGF-1). WFA also induced apoptosis of Ca9-22 cells, which was measured by flow cytometry for subG1 percentage, annexin V expression, and pan-caspase activity, as well as western blotting for caspases 1, 8, and 9 activations. Flow cytometry analysis shows that WFA-treated Ca9-22 oral cancer cells induced G2/M cell cycle arrest, ROS production, mitochondrial membrane depolarization, and phosphorylated histone H2A.X (γH2AX)-based DNA damage. Moreover, pretreating Ca9-22 cells with N-acetylcysteine (NAC) rescued WFA-induced selective killing, apoptosis, G2/M arrest, oxidative stress, and DNA damage. We conclude that WFA induced oxidative stress-mediated selective killing of oral cancer cells.Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We evaluated whether the killing ability of WFA is selective, and we explored its mechanism against oral cancer cells. An MTS tetrazolium cell proliferation assay confirmed that WFA selectively killed two oral cancer cells (Ca9-22 and CAL 27) rather than normal oral cells (HGF-1). WFA also induced apoptosis of Ca9-22 cells, which was measured by flow cytometry for subG1 percentage, annexin V expression, and pan-caspase activity, as well as western blotting for caspases 1, 8, and 9 activations. Flow cytometry analysis shows that WFA-treated Ca9-22 oral cancer cells induced G2/M cell cycle arrest, ROS production, mitochondrial membrane depolarization, and phosphorylated histone H2A.X (γH2AX)-based DNA damage. Moreover, pretreating Ca9-22 cells with N-acetylcysteine (NAC) rescued WFA-induced selective killing, apoptosis, G2/M arrest, oxidative stress, and DNA damage. We conclude that WFA induced oxidative stress-mediated selective killing of oral cancer cells. Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We evaluated whether the killing ability of WFA is selective, and we explored its mechanism against oral cancer cells. An MTS tetrazolium cell proliferation assay confirmed that WFA selectively killed two oral cancer cells (Ca9-22 and CAL 27) rather than normal oral cells (HGF-1). WFA also induced apoptosis of Ca9-22 cells, which was measured by flow cytometry for subG1 percentage, annexin V expression, and pan-caspase activity, as well as western blotting for caspases 1, 8, and 9 activations. Flow cytometry analysis shows that WFA-treated Ca9-22 oral cancer cells induced G2/M cell cycle arrest, ROS production, mitochondrial membrane depolarization, and phosphorylated histone H2A.X (γH2AX)-based DNA damage. Moreover, pretreating Ca9-22 cells with N -acetylcysteine (NAC) rescued WFA-induced selective killing, apoptosis, G2/M arrest, oxidative stress, and DNA damage. We conclude that WFA induced oxidative stress-mediated selective killing of oral cancer cells. Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We evaluated whether the killing ability of WFA is selective, and we explored its mechanism against oral cancer cells. An MTS tetrazolium cell proliferation assay confirmed that WFA selectively killed two oral cancer cells (Ca9-22 and CAL 27) rather than normal oral cells (HGF-1). WFA also induced apoptosis of Ca9-22 cells, which was measured by flow cytometry for subG1 percentage, annexin V expression, and pan-caspase activity, as well as western blotting for caspases 1, 8, and 9 activations. Flow cytometry analysis shows that WFA-treated Ca9-22 oral cancer cells induced G2/M cell cycle arrest, ROS production, mitochondrial membrane depolarization, and phosphorylated histone H2A.X (γH2AX)-based DNA damage. Moreover, pretreating Ca9-22 cells with N-acetylcysteine (NAC) rescued WFA-induced selective killing, apoptosis, G2/M arrest, oxidative stress, and DNA damage. We conclude that WFA induced oxidative stress-mediated selective killing of oral cancer cells. |
| Author | Tang, Jen-Yang Yen, Ching-Yu Huang, Hurng-Wern Chang, Hsueh-Wei Liu, Jing-Ru Li, Ruei-Nian Wang, Hui-Ru Chan, Yu-Hsuan |
| AuthorAffiliation | 1 Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University Kaohsiung, Taiwan 3 Cancer Center, Kaohsiung Medical University Hospital; Kaohsiung Medical University Kaohsiung, Taiwan 6 Institute of Biomedical Science, National Sun Yat-Sen University Kaohsiung, Taiwan 2 Department of Medical Research, Kaohsiung Medical University Hospital Kaohsiung, Taiwan 4 Research Center for Natural Products and Drug Development, Kaohsiung Medical University Kaohsiung, Taiwan 7 Department of Radiation Oncology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University Kaohsiung, Taiwan 5 Institute of Medical Science and Technology, National Sun Yat-Sen University Kaohsiung, Taiwan 9 Department of Radiation Oncology, Kaohsiung Municipal Ta-Tung Hospital Kaohsiung, Taiwan 8 Department of Radiation Oncology, Kaohsiung Medical University Hospital Kaohsiung, Taiwan 10 Department of Oral and Maxillofacial Surgery Chi-Mei Medical Center Tainan, Taiwan 11 School of Dentistry, |
| AuthorAffiliation_xml | – name: 11 School of Dentistry, Taipei Medical University Taipei, Taiwan – name: 4 Research Center for Natural Products and Drug Development, Kaohsiung Medical University Kaohsiung, Taiwan – name: 9 Department of Radiation Oncology, Kaohsiung Municipal Ta-Tung Hospital Kaohsiung, Taiwan – name: 10 Department of Oral and Maxillofacial Surgery Chi-Mei Medical Center Tainan, Taiwan – name: 1 Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University Kaohsiung, Taiwan – name: 3 Cancer Center, Kaohsiung Medical University Hospital; Kaohsiung Medical University Kaohsiung, Taiwan – name: 2 Department of Medical Research, Kaohsiung Medical University Hospital Kaohsiung, Taiwan – name: 7 Department of Radiation Oncology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University Kaohsiung, Taiwan – name: 8 Department of Radiation Oncology, Kaohsiung Medical University Hospital Kaohsiung, Taiwan – name: 6 Institute of Biomedical Science, National Sun Yat-Sen University Kaohsiung, Taiwan – name: 5 Institute of Medical Science and Technology, National Sun Yat-Sen University Kaohsiung, Taiwan |
| Author_xml | – sequence: 1 givenname: Hsueh-Wei surname: Chang fullname: Chang, Hsueh-Wei – sequence: 2 givenname: Ruei-Nian surname: Li fullname: Li, Ruei-Nian – sequence: 3 givenname: Hui-Ru surname: Wang fullname: Wang, Hui-Ru – sequence: 4 givenname: Jing-Ru surname: Liu fullname: Liu, Jing-Ru – sequence: 5 givenname: Jen-Yang surname: Tang fullname: Tang, Jen-Yang – sequence: 6 givenname: Hurng-Wern surname: Huang fullname: Huang, Hurng-Wern – sequence: 7 givenname: Yu-Hsuan surname: Chan fullname: Chan, Yu-Hsuan – sequence: 8 givenname: Ching-Yu surname: Yen fullname: Yen, Ching-Yu |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/28936177$$D View this record in MEDLINE/PubMed |
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| Copyright | Copyright © 2017 Chang, Li, Wang, Liu, Tang, Huang, Chan and Yen. 2017 Chang, Li, Wang, Liu, Tang, Huang, Chan and Yen |
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| Keywords | selective killing apoptosis withaferin A oral cancer oxidative stress N-acetylcysteine |
| Language | English |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Reviewed by: Vivek Choudhary, Augusta University, United States; Paola Patrignani, Università degli Studi “G. d'Annunzio” Chieti-Pescara, Italy; Juan Antonio Rosado Dionisio, University of Extremadura, Spain Edited by: Gareth Davison, Ulster University, United Kingdom This article was submitted to Oxidant Physiology, a section of the journal Frontiers in Physiology |
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| Title | Withaferin A Induces Oxidative Stress-Mediated Apoptosis and DNA Damage in Oral Cancer Cells |
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