Exploring the Functions of Mutant p53 through TP53 Knockout in HaCaT Keratinocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Treatment
2.2. Transcriptomic Analysis
2.3. Proteomic Analysis
2.4. Bioinformatic Analysis
2.5. RT-qPCR
2.6. Migration Assay
2.7. Confocal Fluorescent Microscopy
2.8. Proliferation Assay
2.9. Flow Cytometry
2.10. Statistical Analysis
3. Results
3.1. Transcriptomic Differences
3.2. Proteomic Differences
3.3. p53-Defficient HaCaT Cells Display the Features of EMT
3.4. Knockout of TP53 Enhances Migration and Suppresses Epidermal Differentiation in HaCaT Cells
3.5. p53-Defficient HaCaT keratinocytes Possess Altered PD-L1 Expression
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Forward Primer | Reverse Primer |
---|---|---|
GAPDH | 5′- TCGACAGTCAGCCGCATCTTCTTT -3′ | 5′- ACCAAATCCGTTGACTCCGACCTT -3′ |
ACTB | 5′- TCAGAAGGATTCCTATGTGGGCGA -3′ | 5′- CACGCAGCTCATTGTAGAAGGTGT -3′ |
KRT10 | 5′- AGCATGGCAACTCACATCA -3′ | 5′- GTCGATCTGAAGCAGGATGTT -3′ |
KRT1 | 5′- GCGGACAAATGCAGAGAATG -3′ | 5′- TGCTTGGTAGAGTGCTGTAAG -3′ |
IVL | 5′- 5′-CCAAAGCCTCTGCCTCAG -3′ | 5′- GTATTGACTGGAGGAGGAACAG -3′ |
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Romashin, D.; Rusanov, A.; Arzumanian, V.; Varshaver, A.; Poverennaya, E.; Vakhrushev, I.; Netrusov, A.; Luzgina, N. Exploring the Functions of Mutant p53 through TP53 Knockout in HaCaT Keratinocytes. Curr. Issues Mol. Biol. 2024, 46, 1451-1466. https://doi.org/10.3390/cimb46020094
Romashin D, Rusanov A, Arzumanian V, Varshaver A, Poverennaya E, Vakhrushev I, Netrusov A, Luzgina N. Exploring the Functions of Mutant p53 through TP53 Knockout in HaCaT Keratinocytes. Current Issues in Molecular Biology. 2024; 46(2):1451-1466. https://doi.org/10.3390/cimb46020094
Chicago/Turabian StyleRomashin, Daniil, Alexander Rusanov, Viktoriia Arzumanian, Alexandra Varshaver, Ekaterina Poverennaya, Igor Vakhrushev, Alexander Netrusov, and Nataliya Luzgina. 2024. "Exploring the Functions of Mutant p53 through TP53 Knockout in HaCaT Keratinocytes" Current Issues in Molecular Biology 46, no. 2: 1451-1466. https://doi.org/10.3390/cimb46020094