Inhibition of DNA Repair Pathways and Induction of ROS Are Potential Mechanisms of Action of the Small Molecule Inhibitor BOLD-100 in Breast Cancer
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. BOLD-100 Inhibits Growth of Breast Cancer Cells
2.2. Significantly Altered Genes, Metabolites and Proteins in BOLD-100 Treated Cells Show Changes in DNA Repair Pathways
2.3. BOLD-100 Induces Reactive Oxygen Species (ROS)
2.4. BOLD-100 Synergizes with Anticancer Agents that Target DNA in TNBC Cells
2.5. Combination of BOLD-100 and Olaparib Suppressed Growth of TNBC Tumors
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Reagents
4.2. Cell Proliferation and Viability
4.3. Cell Cycle Analysis
4.4. Analysis of Cellular ROS Assay
4.5. Western Blotting
4.6. Generation, Analysis and Integration of Transcriptomics and Metabolomics Data from MCF7(2) Cells
4.7. Quantification of Citrate/Isocitrate
4.8. TNBC Xenografts and In Vivo Studies
4.9. Immunohistochemistry (IHC)
4.10. Reverse Phase Protein Array (RPPA)
4.11. Statistical Analysis and Drug Interaction for Cell Proliferation Experiments
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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PROTEIN NAME | p-Value | ≥20% Difference in 100 μM BOLD-100 Treated Cells |
---|---|---|
PDK1 | <0.001 | 24% increased |
TSC1 | <0.001 | 27% increased |
SOD2 | <0.001 | 22% increased |
G6PD | <0.001 | 39% increased |
HER2 | 0.002 | 31% increased |
PDK1_pS241 | 0.004 | 35% increased |
Notch1 | 0.006 | 21% increased |
TFAM | 0.006 | 30% increased |
PR (Progesterone receptor) | 0.009 | 64% decreased |
DJ1 | 0.010 | 24% increased |
MUC1 (EMA) | 0.014 | 65% increased |
UGT1A | 0.015 | 36% increased |
Collagen-VI | 0.025 | 21% increased |
c-Jun_pS73 | 0.026 | 20% decreased |
ATRX | 0.031 | 24% decreased |
LC3A-B | 0.035 | 21% increased |
Cyclin-D3 | 0.044 | 39% increased |
Group | n | Administration of BOLD-100 | Administration of Olaparib | ||||||
---|---|---|---|---|---|---|---|---|---|
Drug | Route | mg/kg | Schedule | Drug | Route | mg/kg | Schedule | ||
#1 | 8 | Vehicle | iv | -- | qd4 to end | -- | -- | -- | -- |
#2 | 8 | BOLD-100 | iv | 30 | qd4 to end | -- | -- | -- | -- |
#3 | 8 | BOLD-100 | iv | 50 | qwk to end | -- | -- | -- | -- |
#4 | 8 | olaparib | po | 50 | qd × 32 (start on Day 2) | -- | -- | -- | -- |
#5 | 8 | BOLD-100 | iv | 30 | q4d to end | olaparib | 50 | po | qd × 32 (start on Day 2) |
#6 | 8 | BOLD-100 | iv | 50 | qwk to end | olaparib | 50 | po | qd × 32 (start on Day 2) |
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Bakewell, S.; Conde, I.; Fallah, Y.; McCoy, M.; Jin, L.; Shajahan-Haq, A.N. Inhibition of DNA Repair Pathways and Induction of ROS Are Potential Mechanisms of Action of the Small Molecule Inhibitor BOLD-100 in Breast Cancer. Cancers 2020, 12, 2647. https://doi.org/10.3390/cancers12092647
Bakewell S, Conde I, Fallah Y, McCoy M, Jin L, Shajahan-Haq AN. Inhibition of DNA Repair Pathways and Induction of ROS Are Potential Mechanisms of Action of the Small Molecule Inhibitor BOLD-100 in Breast Cancer. Cancers. 2020; 12(9):2647. https://doi.org/10.3390/cancers12092647
Chicago/Turabian StyleBakewell, Suzanne, Isabel Conde, Yassi Fallah, Mathew McCoy, Lu Jin, and Ayesha N. Shajahan-Haq. 2020. "Inhibition of DNA Repair Pathways and Induction of ROS Are Potential Mechanisms of Action of the Small Molecule Inhibitor BOLD-100 in Breast Cancer" Cancers 12, no. 9: 2647. https://doi.org/10.3390/cancers12092647