Phenanthroindolizidine Alkaloids Isolated from Tylophora ovata as Potent Inhibitors of Inflammation, Spheroid Growth, and Invasion of Triple-Negative Breast Cancer
Abstract
:1. Introduction
2. Results
2.1. Phenanthroindolizidine Alkaloid (PA) Library
2.2. Phenanthroindolizidine Alkaloids (PAs) Are Potent NFκB-Inhibitors in TNBC
2.3. PAs Reduce Cell Viability in the 3D TNBC Co-Culture Model
2.4. Compound (1) Exhibits Superior Anti-Tumor Capacity against TNBC Compared to Paclitaxel
2.5. Compound (1s) Blocks Invasion of TNBC Monoculture 3D Spheroids
2.6. Compound (1s) Blocks HIF-Regulated Transcription
2.7. Compound (1s) Inhibits NFκB Activation Via IκBα Stabilization
2.8. Compound (1s) Blocks Cell Proliferation in the G0/G1 Phase and Delays Cell Cycle Progression
3. Discussion
3.1. PAs Block Inflammation under Hypoxia in TNBC
3.2. PAs Block Proliferation in TNBC through Arrest at the G0/G1-State and Retardation in Cell Cycle Progression
3.3. PAs Maintain Anti-Tumor Potential in a 3D Co-Culture Model Comprising Murine CAFs and Block TNBC Invasion in a 3D Monoculture Spheroid
3.4. Compound (1)/(1s) Exhibits Superior Anti-Tumor Potential Compared to the Chemotherapeutic Agent Paclitaxel
4. Materials and Methods
4.1. Cell Culture and Compounds
4.2. Extraction and Purification of a Phenanthroindolizidines Alkaloids Library
4.3. Chemical Synthesis of O-Methyltylophorinidine (1s)
4.4. Nuclear Factor Kappa B (NFκB) Inhibition Studies
4.4.1. NFκB Inhibition Assay (2 h)
4.4.2. NFκB Inhibition Assay (24 h)
4.5. 2D Cell Viability
4.6. 3D Co-Culture Studies
4.6.1. Spheroid Immunostaining
4.6.2. 3D Cell Viability Studies
4.6.3. 3D Growth Studies
4.7. 3D Invasion Studies
4.8. Hypoxia Studies
4.8.1. HIF Inhibition
4.8.2. Western Blot
4.8.3. Cell Cycle Studies
4.9. Statistical Analysis
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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NFκB Inhibition | Reduction of Cell Viability | ||
---|---|---|---|
Compound | 2 h 1 IC50 [nM] 4 | 24 h 2 IC50 [nM] 4 | 24 h 3 IC50 [nM] 4,5,6 |
(1) | 17.1 ± 2.0 | 3.7 ± 1 | 13.6 ± 0.4 5 |
(2) | 211.8 ± 69.9 | 38.2 ± 14.2 | 117.9 ± 35.0 4 |
(3) | 284.9 ± 60.4 | 114.5 ± 17.9 | 55.3 ± 13.2 4 |
(4) | 83.0 ± 14.7 | 28.3 ± 5.6 | 127 ± 21.4 4 |
(5) | >1000 | >1000 | >1000 6 |
(6) | >1000 | >1000 | >1000 6 |
Paclitaxel | >1000 | >1000 | 37.4 ± 8.6 4 |
Compound | Reduction of Cell Viability 72 h 1 IC50 [nM] 2 |
---|---|
(1) | 21.7 ± 2.5 |
(2) | 441.5 ± 70.8 |
(3) | 476.7 ± 160.4 |
(4) | 517.7 ± 76.8 |
(5) | >1000 |
(6) | >1000 |
Paclitaxel | 43 ± 14.3 |
Compound | Dilution Factor | Concentration Range |
---|---|---|
(1), (2), (3), (4), (5), (6) | 4 | 1000 nM to 0.06 nM |
(1s) | 3 | 100 nM to 0.05 nM |
Paclitaxel | 3 | 1000 nM to 0.5 nM |
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Reimche, I.; Yu, H.; Ariantari, N.P.; Liu, Z.; Merkens, K.; Rotfuß, S.; Peter, K.; Jungwirth, U.; Bauer, N.; Kiefer, F.; et al. Phenanthroindolizidine Alkaloids Isolated from Tylophora ovata as Potent Inhibitors of Inflammation, Spheroid Growth, and Invasion of Triple-Negative Breast Cancer. Int. J. Mol. Sci. 2022, 23, 10319. https://doi.org/10.3390/ijms231810319
Reimche I, Yu H, Ariantari NP, Liu Z, Merkens K, Rotfuß S, Peter K, Jungwirth U, Bauer N, Kiefer F, et al. Phenanthroindolizidine Alkaloids Isolated from Tylophora ovata as Potent Inhibitors of Inflammation, Spheroid Growth, and Invasion of Triple-Negative Breast Cancer. International Journal of Molecular Sciences. 2022; 23(18):10319. https://doi.org/10.3390/ijms231810319
Chicago/Turabian StyleReimche, Irene, Haiqian Yu, Ni Putu Ariantari, Zhen Liu, Kay Merkens, Stella Rotfuß, Karin Peter, Ute Jungwirth, Nadine Bauer, Friedemann Kiefer, and et al. 2022. "Phenanthroindolizidine Alkaloids Isolated from Tylophora ovata as Potent Inhibitors of Inflammation, Spheroid Growth, and Invasion of Triple-Negative Breast Cancer" International Journal of Molecular Sciences 23, no. 18: 10319. https://doi.org/10.3390/ijms231810319