Upregulation of CASP9 through NF-κB and Its Target MiR-1276 Contributed to TNFα-Promoted Apoptosis of Cancer Cells Induced by Doxorubicin
Abstract
:1. Introduction
2. Results
2.1. NF-κB Binds to CASP9
2.2. NF-κB Directly Regulates CASP9 Expression
2.3. NF-κB Indirectly Regulates CASP9 Expression
2.4. The Regulation of CASP9 by NF-κB-mediated FFL Contributes to the TNFα Promotion of DOX-Induced Apoptosis
3. Discussion
4. Materials and Methods
4.1. ChIP-Seq Data Analysis
4.2. Cell Culture and Treatment
4.3. ChIP Expriment
4.4. ChIP-qPCR
4.5. Luciferase Reporter Assay
4.6. Cell Viability and Apoptosis Detection
4.7. Real-time Reverse Transcription-PCR
4.8. Western Blotassay
4.9. miRNA Target Prediction
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
BSs | NF-κB-binding sites |
CASP | Caspase |
cCASP9 | cleaved CASP9 |
DOX | doxorubicin |
fCASP9 | full-length CASP9 protein |
TF | transcription factor |
FFL | feed-forward loop |
RQ | relative quantification |
tCASP9 | total CASP9 |
ChIP-Seq | Chromatin Immunoprecipitation-Sequencing |
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BSs | FE | κB Number | Location |
---|---|---|---|
BS1 | 20.579 | 44 | 3′ region |
BS2 | 21.720 | 23 | 3′ region |
BS3 | 21.236 | 55 | 3′ region |
BS4 | 34.436 | 14 | 3′ region |
BS5 | 30.491 | 8 | 3′ region |
BS6 | 21.982 | 8 | 3′ region |
BS7 | 22.187 | 21 | 3′ region |
BS8 | 21.805 | 3 | 3′ region |
BS9 | 20.638 | 31 | 3′ region |
BS10 | 27.507 | 17 | 5′ region |
BS11 | 21.918 | 19 | 5′ region |
Average | 24.045 | 22.091 |
BSs | FE | κB Number | Location |
---|---|---|---|
BS1 | 22.87 | 3 | 3′ region |
BS2 | 21.869 | 4 | 3′ region |
BS3 | 20.815 | 10 | 3′ region |
BS4 | 32.358 | 28 | 3′ region |
BS5 | 20.386 | 68 | 3′ region |
BS6 | 21.175 | 77 | Internal |
BS7 | 25.373 | 55 | Internal |
BS8 | 23.653 | 30 | Internal |
BS9 | 28.99 | 35 | 5′ region |
BS10 | 25.738 | 4 | 5′ region |
BS11 | 29.767 | 2 | 5′ region |
Average | 24.818 | 28.727 |
miRWalk | TargetScan | DIANA-microT-CDS | |
---|---|---|---|
Binding type | 7 mer | 7 mer-1A/7 mer-m8 | 7 mer/6 mer/6 mer/7 mer/6 mer/6 mer/7 mer(CDS) |
Potential binding site | 1 | 1 | 7 (6 in 3′ UTR +1 in CDS) |
Region | 3′ UTR | 3′ UTR | 3′ UTR and CDS |
Confidence | P value = 0.041 | Context + +score = −0.11 | miTG core = 0.766 |
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Zhou, F.; Li, Y.; Huang, Y.; Wu, J.; Wu, Q.; Zhu, H.; Wang, J. Upregulation of CASP9 through NF-κB and Its Target MiR-1276 Contributed to TNFα-Promoted Apoptosis of Cancer Cells Induced by Doxorubicin. Int. J. Mol. Sci. 2020, 21, 2290. https://doi.org/10.3390/ijms21072290
Zhou F, Li Y, Huang Y, Wu J, Wu Q, Zhu H, Wang J. Upregulation of CASP9 through NF-κB and Its Target MiR-1276 Contributed to TNFα-Promoted Apoptosis of Cancer Cells Induced by Doxorubicin. International Journal of Molecular Sciences. 2020; 21(7):2290. https://doi.org/10.3390/ijms21072290
Chicago/Turabian StyleZhou, Fei, Yun Li, Yisheng Huang, Jian Wu, Qinhan Wu, Hui Zhu, and Jinke Wang. 2020. "Upregulation of CASP9 through NF-κB and Its Target MiR-1276 Contributed to TNFα-Promoted Apoptosis of Cancer Cells Induced by Doxorubicin" International Journal of Molecular Sciences 21, no. 7: 2290. https://doi.org/10.3390/ijms21072290