Identification of TAT as a Biomarker Involved in Cell Cycle and DNA Repair in Breast Cancer
Abstract
:1. Introduction
2. Materials and Methods
2.1. The Analysis for Differential Gene Expression and Promoter Methylation Level
2.2. Receiver Operating Characteristic (ROC) Curve
2.3. Kaplan–Meier Plotter (KM Plotter)
2.4. Human Protein Atlas (HPA)
2.5. Correlation Analysis of Gene Expression
2.6. Gene Expression Omnibus (GEO)
2.7. Gene Set Enrichment Analysis (GSEA)
2.8. Target Overlap Analysis
2.9. Molecular Docking
2.10. Cell Culture
2.11. RNA Extraction and Real-Time PCR Analysis (qPCR)
2.12. Cell Proliferation Assay
2.13. Western Blot
2.14. Colony Formation Assay
2.15. Reagent and Antibodies (Table 2)
Reagent or Antibody | Source | Identifier |
---|---|---|
Diazinon | TargetMol | T0998 |
TAT | Biodragon | BD-PE0222 |
DNMT3A | Selleck | F1036 |
DNMT3B | Selleck | F0001 |
γH2Ax | ABclonal | AP0687 |
CyclinD1 | proteintech | 60186-1-Ig |
CDK4 | proteintech | 11026-1-AP |
Actin | proteintech | 66009-1-Ig |
2.16. Statistics
3. Results
3.1. TAT Was Down-Regulated and Associated with Poor Prognosis in BC
3.2. TAT Promoter Was Methylated by DNMT3A and DNMT3B
3.3. DNMT3A and DNMT3B Reduce TAT Expression by Methylation of Its Promoter in Triple-Negative Breast Cancer
3.4. Reduced TAT Contributes to Activation of the Cell Cycle and Promotes DNA Repair
3.5. Targeting DNMT3A/3B Inhibits Proliferation of Breast Cancer Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene | Sequence (5′→3′) |
---|---|
TAT | Primer-F: CTGGACTCGGGCAAATATAATGG Primer-R: GTCCTTAGCTTCTAGGGGTGC |
CDC6 | Primer-F: CCAGGCACAGGCTACAATCAG Primer-R: AACAGGTTACGGTTTGGACATT |
RPA3 | Primer-F: AGCTCAATTCATCGACAAGCC Primer-R: TCTTCATCAAGGGGTTCCATCA |
POLD1 | Primer-F: CAGTGCCAAGGTGGTGTATGG Primer-R: CTTGCTGATAAGCAGGTATGGG |
POLD2 | Primer-F: CAGCGTATCAAACTAAAAGGCAC Primer-R: GTCTCTCACGGAGCCAAACAC |
ACTIN | Primer-F: GGAAATCGTGCGTGACAT Primer-R: TGCCAATGGTGATGACCT |
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Xie, F.; Hua, S.; Guo, Y.; Wang, T.; Shan, C.; Zhang, L.; He, T. Identification of TAT as a Biomarker Involved in Cell Cycle and DNA Repair in Breast Cancer. Biomolecules 2024, 14, 1088. https://doi.org/10.3390/biom14091088
Xie F, Hua S, Guo Y, Wang T, Shan C, Zhang L, He T. Identification of TAT as a Biomarker Involved in Cell Cycle and DNA Repair in Breast Cancer. Biomolecules. 2024; 14(9):1088. https://doi.org/10.3390/biom14091088
Chicago/Turabian StyleXie, Fei, Saiwei Hua, Yajuan Guo, Taoyuan Wang, Changliang Shan, Lianwen Zhang, and Tao He. 2024. "Identification of TAT as a Biomarker Involved in Cell Cycle and DNA Repair in Breast Cancer" Biomolecules 14, no. 9: 1088. https://doi.org/10.3390/biom14091088