FANCD2 Confers a Malignant Phenotype in Esophageal Squamous Cell Carcinoma by Regulating Cell Cycle Progression
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Clinical Specimens
2.2. RNA Sequence Analysis
2.3. Cell Lines
2.4. Plasmids and Lentivirus Preparation and Infection
2.5. Western Blot Analysis
2.6. In Vivo Tumorigenicity Assay
2.7. MTT Assay
2.8. Colony Formation Assay
2.9. Synchronization and Flow Cytometry Analysis
2.10. In Vitro Chemotherapy Treatment
2.11. Subcellular Fractionation
2.12. Statistical Analysis
3. Results
3.1. Upregulation of FANCD2 Gene Expression in ESCC Tissues and Cell Lines
3.2. Depletion of FANCD2 Protein Expression Inhibits In Vivo Tumor Growth and Metastasis
3.3. FANCD2-KO Suppresses In Vitro Cell Proliferation and Colony Formation in ESCC Cells
3.4. FANCD2-KO Suppresses Cell Cycle Progression in ESCC Cells
3.5. FANCD2-KO Affects G1/S Transition and Delays Entry to Mitosis in ESCC Cells
3.6. Ubiquitinated FANCD2 Localizes to the Nucleus to Regulate Cell Cycle Progression in ESCC Cells
3.7. FANCD2 Is Likely Dispensable for DNA Damage Repair in ESCC Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Antibody | Company | Catalog Number | Dilution | Antibody | Company | Catalog Number | Dilution |
---|---|---|---|---|---|---|---|
FANCD2 | Santa Cruz | sc20022 | 1:1000 | CDK4 | Cell Signaling | 12,790 | 1:1000 |
p84 | GeneTex | GTX70220 | 1:2000 | CDK7 | Cell Signaling | 2090 | 1:1000 |
Cyclin A2 | Cell Signaling | 4656 | 1:1000 | CDK9 | Cell Signaling | 2316 | 1:1000 |
Cyclin B1 | Cell Signaling | 4138 | 1:1000 | p-ATR | Cell Signaling | 2853 | 1:1000 |
Cyclin D1 | Cell Signaling | 2978 | 1:1000 | p-ATM | Cell Signaling | 13,050 | 1:1000 |
Cyclin D2 | Cell Signaling | 3741 | 1:1000 | p-Chk1 | Cell Signaling | 12,302 | 1:1000 |
Cyclin E2 | Cell Signaling | 4132 | 1:1000 | p-Chk2 | Cell Signaling | 2661 | 1:1000 |
Chk1 | Cell Signaling | 2345 | 1:500 | p-BRCA1 | Cell Signaling | 9009 | 1:1000 |
Chk2 | Cell Signaling | 3440 | 1:1000 | Vinculin | Cell Signaling | 13,901 | 1:2000 |
cdc2 | Cell Signaling | 28,439 | 1:1000 | ⍺-Tubulin | Cell Signaling | 2125 | 1:2000 |
CDK2 | Cell Signaling | 2546 | 1:1000 |
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Lei, L.C.; Yu, V.Z.; Ko, J.M.Y.; Ning, L.; Lung, M.L. FANCD2 Confers a Malignant Phenotype in Esophageal Squamous Cell Carcinoma by Regulating Cell Cycle Progression. Cancers 2020, 12, 2545. https://doi.org/10.3390/cancers12092545
Lei LC, Yu VZ, Ko JMY, Ning L, Lung ML. FANCD2 Confers a Malignant Phenotype in Esophageal Squamous Cell Carcinoma by Regulating Cell Cycle Progression. Cancers. 2020; 12(9):2545. https://doi.org/10.3390/cancers12092545
Chicago/Turabian StyleLei, Lisa Chan, Valen Zhuoyou Yu, Josephine Mun Yee Ko, Lvwen Ning, and Maria Li Lung. 2020. "FANCD2 Confers a Malignant Phenotype in Esophageal Squamous Cell Carcinoma by Regulating Cell Cycle Progression" Cancers 12, no. 9: 2545. https://doi.org/10.3390/cancers12092545