ASPP2 Is Phosphorylated by CDK1 during Mitosis and Required for Pancreatic Cancer Cell Proliferation
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Culture and Transfection, and Transduction
2.2. Anti-Mitotic Drugs and Transfection Agents
2.3. Expression Constructs and RNA Interference
- shASPP2-A: GTCTAGTAAATAGGATCATTT.
- shASPP2-B: GAAATCCAGAATCCATATTTA.
2.4. Phos-Tag, Western Blotting, and Peptide Blocking
2.5. Recombinant Protein Purification and In Vitro Kinase Assay
2.6. Antibodies
2.7. Cell Proliferation
2.8. Immunofluorescence Staining and Confocal Microscopy
2.9. RNA Isolation, Reverse Transcription, and RT-PCR
2.10. RNA-Seq
2.11. Xenograft Mouse Model
2.12. Statistical Analysis
3. Results
3.1. The ASPP Family of Proteins Are Phosphorylated during Mitosis
3.2. ASPP2 Is Phosphorylated at S562 and S704 by CDK1 during Mitosis
3.3. ASPP2 Is Overexpressed and Required for Pancreatic Cancer Growth
3.4. ASPP2 Regulates the Expression of YAP-Associated Genes
3.5. RNA-Seq Identified Genes Correlating with ASPP2 Expression and Implying Negative Patient Outcome
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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Xiao, Y.; Chen, Y.; Chen, J.; Dong, J. ASPP2 Is Phosphorylated by CDK1 during Mitosis and Required for Pancreatic Cancer Cell Proliferation. Cancers 2023, 15, 5424. https://doi.org/10.3390/cancers15225424
Xiao Y, Chen Y, Chen J, Dong J. ASPP2 Is Phosphorylated by CDK1 during Mitosis and Required for Pancreatic Cancer Cell Proliferation. Cancers. 2023; 15(22):5424. https://doi.org/10.3390/cancers15225424
Chicago/Turabian StyleXiao, Yi, Yuanhong Chen, Jianan Chen, and Jixin Dong. 2023. "ASPP2 Is Phosphorylated by CDK1 during Mitosis and Required for Pancreatic Cancer Cell Proliferation" Cancers 15, no. 22: 5424. https://doi.org/10.3390/cancers15225424