HSP90α Mediates Sorafenib Resistance in Human Hepatocellular Carcinoma by Necroptosis Inhibition under Hypoxia
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. Sorafenib Induced HCC Necroptosis
2.2. Hypoxia Contributed to HCC Resistance to Sorafenib
2.3. Hypoxia Impeded the Distribution of RIPK1/RIPK3/MLKL Complex in Cytoplasm
2.4. HSP90α Promotes Chaperone-Mediated Autophagy (CMA) Degradation by Directly Binding to MLKL in Hypoxia
2.5. 17-AAG Combining with Sorafenib Enhanced Necroptosis Pathway In Vitro
2.6. HSP90α Could Be an Important Target in Sorafenib Resistance In Vivo
2.7. Clinical Analysis of HIF1α/HSP90α as a Therapeutic Target
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture
4.3. Clinical Samples
4.4. PDX (Patient-Derived Tumor Xenograft) Model and Sorafenib—Resistance Model In Vivo
4.5. Animal Studies
4.6. Western Blot Analysis
4.7. Immunohistochemistry (IHC)
4.8. Immunofluorescence Assay
4.9. Cell Transfection
4.10. Cytoplasmic and Nuclear Protein Extraction
4.11. Co-Immunoprecipitation (Co-IP)
4.12. Flowcytometry Analysis
4.13. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Ethics Approval and Consent to Participate
Abbreviations
HCC | Hepatocellular carcinoma |
17-AAG | Demethoxygeldanamycin |
IF | Immunofluorescence |
WB | Western Blot |
IHC | Immunohistochemistry |
i.g. | intragastrically |
Co-IP | Coimmunoprecipitation |
CMA | chaperone-mediated autophagy |
DAB | 3, 3′-diaminobenzidine |
DAPI | 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride |
PI | Propidium Iodide |
PBS | Phosphate-Buffered Saline |
MTT | 3- (4, 5-Dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide |
DMSO | Dimethyl sulfoxide |
Nec | necrostatin-1 |
VAD | Z-VAD-FMK |
RIPK1 | Receptor-interacting protein kinase 1 |
MLKL | Mixed lineage kinase domain-like |
RIPK3 | Receptor-interacting protein kinase-3 |
HSP90α | Heat shock proteins 90α |
HIF1α | Hypoxia inducible factor-1α |
AFP | Alpha fetoprotein |
LAMP2 | Lysosomal-associated membrane protein 2 |
VEGF-R | Vascular endothelial growth factor receptors |
PDGF-R | Platelet-derived growth factor receptor |
PDX | Patient-Derived tumor Xenograft |
GEPIA | Gene Expression Profiling Interactive Analysis |
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Liao, Y.; Yang, Y.; Pan, D.; Ding, Y.; Zhang, H.; Ye, Y.; Li, J.; Zhao, L. HSP90α Mediates Sorafenib Resistance in Human Hepatocellular Carcinoma by Necroptosis Inhibition under Hypoxia. Cancers 2021, 13, 243. https://doi.org/10.3390/cancers13020243
Liao Y, Yang Y, Pan D, Ding Y, Zhang H, Ye Y, Li J, Zhao L. HSP90α Mediates Sorafenib Resistance in Human Hepatocellular Carcinoma by Necroptosis Inhibition under Hypoxia. Cancers. 2021; 13(2):243. https://doi.org/10.3390/cancers13020243
Chicago/Turabian StyleLiao, Yan, Yue Yang, Di Pan, Youxiang Ding, Heng Zhang, Yuting Ye, Jia Li, and Li Zhao. 2021. "HSP90α Mediates Sorafenib Resistance in Human Hepatocellular Carcinoma by Necroptosis Inhibition under Hypoxia" Cancers 13, no. 2: 243. https://doi.org/10.3390/cancers13020243