Elucidating Cuproptosis-Associated Genes in the Progression from Nash to HCC Using Bulk and Single-Cell RNA Sequencing Analyses and Experimental Validation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Data Collection
2.2. Gene Set Enrichment Analysis (GSEA) and OS Analysis of CRGs
2.3. Construction of Nomogram Model for CRGs
2.4. Construction and Verification of CRC for NASH and NASH Development to HCC
2.5. Assessment of Immune Cell Infiltration CRC Using ssGSEA
2.6. Data Processing and Analysis of Single Cells
2.7. Weighted Gene Co-Expression Network Analysis (WGCNA)
2.8. Human Tissue Specimens
2.9. Reagents, Animals, and Animal Experiments
2.10. Biochemical Analysis
2.11. Immunohistochemistry
2.12. Oil Red O Staining
2.13. Histology Examination
2.14. Statistics
3. Results
3.1. Identification of Key CRGs in the Transition of NASH to HCC
3.2. Analysis of the Immune Microenvironment of NASH to HCC
3.3. Construction of Nomogram Models Based on the Development of NASH to HCC
3.4. Construction, Validation, and Biological Characterization of a Cuproptosis-Related Cluster
3.5. Functional Analyses Based on the NASH CRC to HCC CRC and PPI Construction
3.6. Single-Cell Analysis of Healthy Mouse Hepatocytes and Mouse NASH Hepatocytes
3.7. Validation of FDX1 Expression in NASH Models and Human NASH Samples
3.8. GSEA Evaluation Based on FDX1 Expression
4. Discussion
Limitations
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Zhao, Z.; Luan, T.; Wan, J.; Du, H.; Hu, J.; Liu, H.; Gong, X.; Kuang, G.; Wang, B. Elucidating Cuproptosis-Associated Genes in the Progression from Nash to HCC Using Bulk and Single-Cell RNA Sequencing Analyses and Experimental Validation. Medicina 2023, 59, 1639. https://doi.org/10.3390/medicina59091639
Zhao Z, Luan T, Wan J, Du H, Hu J, Liu H, Gong X, Kuang G, Wang B. Elucidating Cuproptosis-Associated Genes in the Progression from Nash to HCC Using Bulk and Single-Cell RNA Sequencing Analyses and Experimental Validation. Medicina. 2023; 59(9):1639. https://doi.org/10.3390/medicina59091639
Chicago/Turabian StyleZhao, Zizuo, Tiankuo Luan, Jingyuan Wan, Hui Du, Jun Hu, Hao Liu, Xia Gong, Ge Kuang, and Bin Wang. 2023. "Elucidating Cuproptosis-Associated Genes in the Progression from Nash to HCC Using Bulk and Single-Cell RNA Sequencing Analyses and Experimental Validation" Medicina 59, no. 9: 1639. https://doi.org/10.3390/medicina59091639