Androgen Receptor-Activated Enhancers Simultaneously Regulate Oncogene TMPRSS2 and lncRNA PRCAT38 in Prostate Cancer
Abstract
:1. Introduction
2. Material and Methods
2.1. Cell Culture
2.2. Cell Proliferation Assay
2.3. RNA Extraction, RT-PCR and Quantitative PCR
2.4. Western Blot Assay
2.5. ChIP-seq and GRO-seq Data Analyses
2.6. RNA Interference (RNAi) Assay
2.7. Chromatin Immunoprecipitation (ChIP)
2.8. Dual-luciferase Reporter Assay
2.9. Knockout of Enhancer Regions
2.10. Chromosome Conformation Capture (3C) Assay
2.11. Cell Migration Assay
2.12. Statistical Analysis
3. Results
3.1. PRCAT38 is an Androgen-Regulated lncRNA that Modulates Cell Growth and Migration in Prostate Cancer
3.2. AR binds to Enhancers between TMPRSS2 and PRCAT38, to Recruit RNA Pol II and H3K27ac to Initiate Transcription
3.3. Enhancer E1 is Responsive to Androgen Induction and Regulates both TMPRSS2 and PRCAT38 via Interacting with E2 through Chromatin Looping
3.4. FOXA1 is Recruited by the AR to the Enhancers and Regulates the Expression of TMPRSS2 and PRCAT38
3.5. FOXA1 is Required for Enhancer Activity and Chromatin Looping between Enhancers and Promoters of TMPRSS2 and PRCAT38
3.6. p300 Inhibition Impedes Enhancer-FOXA1 Binding and Chromatin Looping
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AR | androgen receptor |
Eth | ethanol |
DHT | dihydrotestosterone |
DMSO | dimethylsulfoxide |
TMPRSS2 | transmembrane protease serine 2 |
ETS | E26 transformation-specific |
PRCAT38 | prostate cancer associated transcript 38 |
FOXA1 | forkhead box protein A1 |
Pol II | RNA polymerase II |
SD | standard deviation |
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Chen, Z.; Song, X.; Li, Q.; Xie, L.; Guo, T.; Su, T.; Tang, C.; Chang, X.; Liang, B.; Huang, D. Androgen Receptor-Activated Enhancers Simultaneously Regulate Oncogene TMPRSS2 and lncRNA PRCAT38 in Prostate Cancer. Cells 2019, 8, 864. https://doi.org/10.3390/cells8080864
Chen Z, Song X, Li Q, Xie L, Guo T, Su T, Tang C, Chang X, Liang B, Huang D. Androgen Receptor-Activated Enhancers Simultaneously Regulate Oncogene TMPRSS2 and lncRNA PRCAT38 in Prostate Cancer. Cells. 2019; 8(8):864. https://doi.org/10.3390/cells8080864
Chicago/Turabian StyleChen, Zikai, Xuhong Song, Qidong Li, Lingzhu Xie, Tangfei Guo, Ting Su, Chang Tang, Xiaolan Chang, Bin Liang, and Dongyang Huang. 2019. "Androgen Receptor-Activated Enhancers Simultaneously Regulate Oncogene TMPRSS2 and lncRNA PRCAT38 in Prostate Cancer" Cells 8, no. 8: 864. https://doi.org/10.3390/cells8080864