Expression of SnoRNA U50A Is Associated with Better Prognosis and Prolonged Mitosis in Breast Cancer
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Patient Specimens
2.2. Total RNA Extraction
2.3. RNA-Sequencing Analysis
2.4. Reverse Transcription
2.5. SnoRNA Detection and Absolute/Relative Real-Time PCR
2.6. Plasmid and Transfection
2.7. SnoRNA Inhibition
2.8. Lentiviral Knockdown
2.9. Western Blot
2.10. Immunofluorescence
2.11. In Situ Hybridization (ISH) Detection
2.12. Statistical Analysis
2.13. Colony Formation
3. Results
3.1. U50A Expression Level Is Associated with Better Prognosis in Breast Cancer Patients
3.2. Mitosis-Related Transcriptomic Changes Regulated by U50A
3.3. Identification of the Downstream Targets of U50A Involved in Mitosis
3.4. U50A Prolongs Mitosis and Suppresses Colony-Forming Ability in Breast Cancer Cells
3.5. Correlation of U50A Expression with Mitotic Cells in Human Breast Cancer Tissues
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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Characteristics | No. of Patients (%) |
---|---|
Median age (range, years) | 50 (31–84) |
Age ≤ 50 | 52 (46%) |
Age > 50 | 62 (54%) |
Pathologic characteristic | - |
Invasive ductal carcinoma | 97 (85%) |
Invasive lobular carcinoma | 1 (1%) |
Others | 16 (14%) |
Others: Mucinous carcinoma, papillary carcinoma, and invasive carcinoma | |
Breast cancer subtype | - |
Luminal A/B | 68 (60%) |
HER2-enriched | 28 (24.6%) |
Triple-negative | 18 (15.4%) |
Variable | All Breast Cancer Patients in This Study | |||
---|---|---|---|---|
Overall Survival (OS) | Relapse-Free Survival (RFS) | |||
p | HR | p | HR | |
U50A | 0.02 * | 0.45 # | 0.01 * | 0.43 # |
U50B | 0.32 | 0.7 | 0.36 | 0.72 |
SNHG5 | 0.2 | 0.64 | 0.22 | 0.65 |
All | Stage (Mean ± S.D.) | ||||
---|---|---|---|---|---|
Marker | I | II | III | IV | p |
U50A | 316.3 ± 201.7 | 122.3 ± 98.98 | 177.1 ± 155.3 | 42.2 ± 31.4 | *** <0.0001 |
U50B | 10.48 ± 17.28 | 6.38 ± 10.86 | 5.005 ± 7.4 | 1.705 ± 2.39 | 0.62 |
SNHG5 | 36.04 ± 44.1 | 19.07 ± 38.69 | 60.72 ± 117.1 | 0.49 ± 0.67 | 0.07 |
Selected Targets | Fold Change from RNA-Sequencing (pLUX-U50A/pLUX) |
---|---|
CENPE | 0.49 |
CENPF | 0.58 |
SMC5 | 0.63 |
ATRX | 0.65 |
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Li, J.-N.; Wang, M.-Y.; Chen, Y.-T.; Kuo, Y.-L.; Chen, P.-S. Expression of SnoRNA U50A Is Associated with Better Prognosis and Prolonged Mitosis in Breast Cancer. Cancers 2021, 13, 6304. https://doi.org/10.3390/cancers13246304
Li J-N, Wang M-Y, Chen Y-T, Kuo Y-L, Chen P-S. Expression of SnoRNA U50A Is Associated with Better Prognosis and Prolonged Mitosis in Breast Cancer. Cancers. 2021; 13(24):6304. https://doi.org/10.3390/cancers13246304
Chicago/Turabian StyleLi, Jie-Ning, Ming-Yang Wang, Yi-Ting Chen, Yao-Lung Kuo, and Pai-Sheng Chen. 2021. "Expression of SnoRNA U50A Is Associated with Better Prognosis and Prolonged Mitosis in Breast Cancer" Cancers 13, no. 24: 6304. https://doi.org/10.3390/cancers13246304