Hydroxychloroquine (HCQ) Modulates Autophagy and Oxidative DNA Damage Stress in Hepatocellular Carcinoma to Overcome Sorafenib Resistance via TLR9/SOD1/hsa-miR-30a-5p/Beclin-1 Axis
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Lines and Media
2.2. Microarray Preprocessing and Analysis
2.3. Differential Expression Analysis
2.4. Immunohistochemistry (IHC) Analyses
2.5. Cell Viability Test and Calculation of the Combination Index
2.6. Apoptosis and Cell Cycle Analysis
2.7. Western Blotting and qRT-PCR
2.8. Colony Formation Assay
2.9. Wound Healing Migration Assay
2.10. Invasion Assay
2.11. Sphere Formation Assay
2.12. Animal Studies
2.13. Statistical Analysis
3. Results
3.1. TLR9 Overexpressed in HCC Sorafenib-Resistant HCC Tissue and Cells
3.2. Hydroxychloroquine (HCQ) Suppressed the Cell Proliferation and Viability of HCC Cell Lines
3.3. HCQ Treatment Inhibited Tumor Spheroid Growth and Stem/Self-Renewal Properties of HCC Cell Lines
3.4. HCQ Inhibited Oncogenic Potential by Reducing Migration and Invasion of HCC Cells
3.5. HCQ Modulated the Expression of Oxidative Stress Marker (SOD1) in HCC Cells
3.6. HCQ Displayed a Synergistic Effect with the Sorafenib on HCC Sorafenib-Resistant Cells
3.7. HCQ Treatment Modulated the Autophagy and Apoptosis of HCC Sorafenib-Resistant Cells In Vitro
3.8. MiR-30a-5p Regulated the Expression of Autophagy Cell Markers in Sorafenib-Resistant HCC Cells
3.9. In Vivo Effect of HCQ Treatment Sensitized Sorafenib Efficacy on Sorafenib-Resistant HCC Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Chen, M.-Y.; Yadav, V.K.; Chu, Y.C.; Ong, J.R.; Huang, T.-Y.; Lee, K.-F.; Lee, K.-H.; Yeh, C.-T.; Lee, W.-H. Hydroxychloroquine (HCQ) Modulates Autophagy and Oxidative DNA Damage Stress in Hepatocellular Carcinoma to Overcome Sorafenib Resistance via TLR9/SOD1/hsa-miR-30a-5p/Beclin-1 Axis. Cancers 2021, 13, 3227. https://doi.org/10.3390/cancers13133227
Chen M-Y, Yadav VK, Chu YC, Ong JR, Huang T-Y, Lee K-F, Lee K-H, Yeh C-T, Lee W-H. Hydroxychloroquine (HCQ) Modulates Autophagy and Oxidative DNA Damage Stress in Hepatocellular Carcinoma to Overcome Sorafenib Resistance via TLR9/SOD1/hsa-miR-30a-5p/Beclin-1 Axis. Cancers. 2021; 13(13):3227. https://doi.org/10.3390/cancers13133227
Chicago/Turabian StyleChen, Ming-Yao, Vijesh Kumar Yadav, Yi Cheng Chu, Jiann Ruey Ong, Ting-Yi Huang, Kwai-Fong Lee, Kuen-Haur Lee, Chi-Tai Yeh, and Wei-Hwa Lee. 2021. "Hydroxychloroquine (HCQ) Modulates Autophagy and Oxidative DNA Damage Stress in Hepatocellular Carcinoma to Overcome Sorafenib Resistance via TLR9/SOD1/hsa-miR-30a-5p/Beclin-1 Axis" Cancers 13, no. 13: 3227. https://doi.org/10.3390/cancers13133227