AF8c, a Multi-Kinase Inhibitor Induces Apoptosis by Activating DR5/Nrf2 via ROS in Colorectal Cancer Cells
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cells and Cell Culture
2.2. Chemicals and Antibodies
2.3. Cell Viability Assay
2.4. Colony Formation
2.5. Western Blot Analysis
2.6. Analysis of Apoptosis
2.7. Determination of Mitochondrial ROS
2.8. Transfection
2.9. RNA Isolation, and Quantitative Real-Time PCR (qRT-PCR)
2.10. Immunoprecipitation (IP)
2.11. Fraction
2.12. Immunofluorescence Staining
2.13. 3D Cell Culture
2.14. In Vivo Tumor Xenograft Study
2.15. Immunohistochemistry
2.16. Terminal Deoxyribonucleotidyl Transferase-Mediated Deoxyuridine Triphosphate Nick-End Labeling (TUNEL) Assay
2.17. Patient-Derived Colorectal Cancer (PDC) Cells
2.18. Statistical Analysis
3. Results
3.1. Treatment with AF8c Inhibits Cell Viability and Induces Apoptosis in Human CRC Cells
3.2. AF8c-Mediated Upregulation of DR5 Is a Factor for Apoptosis Induction
3.3. ER Stress and ROS Leads to Apoptosis in Cells Exposed to AF8c
3.4. Activation of DR5-Induced Nrf2 by AF8c Is a Prerequisite for Apoptotic Cell Death
3.5. AF8c Enhances Apoptotic Cell Death in PDC Cells through Activation of DR5 and Nrf2 by Increased ER Stress
3.6. AF8c Suppresses Tumor Growth In Vivo by Increasing DR5, Nrf2, and CHOP Expression and Subsequent Apoptosis
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Jeong, S.; Farag, A.K.; Yun, H.K.; Jeong, Y.A.; Kim, D.Y.; Jo, M.J.; Park, S.H.; Kim, B.R.; Kim, J.L.; Kim, B.G.; et al. AF8c, a Multi-Kinase Inhibitor Induces Apoptosis by Activating DR5/Nrf2 via ROS in Colorectal Cancer Cells. Cancers 2022, 14, 3043. https://doi.org/10.3390/cancers14133043
Jeong S, Farag AK, Yun HK, Jeong YA, Kim DY, Jo MJ, Park SH, Kim BR, Kim JL, Kim BG, et al. AF8c, a Multi-Kinase Inhibitor Induces Apoptosis by Activating DR5/Nrf2 via ROS in Colorectal Cancer Cells. Cancers. 2022; 14(13):3043. https://doi.org/10.3390/cancers14133043
Chicago/Turabian StyleJeong, Soyeon, Ahmed K. Farag, Hye Kyeong Yun, Yoon A. Jeong, Dae Yeong Kim, Min Jee Jo, Seong Hye Park, Bo Ram Kim, Jung Lim Kim, Bu Gyeom Kim, and et al. 2022. "AF8c, a Multi-Kinase Inhibitor Induces Apoptosis by Activating DR5/Nrf2 via ROS in Colorectal Cancer Cells" Cancers 14, no. 13: 3043. https://doi.org/10.3390/cancers14133043