Cytotoxic and Antiproliferative Effects of Preussin, a Hydroxypyrrolidine Derivative from the Marine Sponge-Associated Fungus Aspergillus candidus KUFA 0062, in a Panel of Breast Cancer Cell Lines and Using 2D and 3D Cultures
Abstract
:1. Introduction
2. Results
2.1. Cells Exposure in 2D
2.1.1. Analysis of Cell Viability—MTT Assay
2.1.2. Analysis of Cell Proliferation—5-bromo-2′-deoxyuridine (BrdU) Assay
2.1.3. Cell Morphology
2.2. Cells Exposure in 3D
2.2.1. Analysis of Cell Viability
MTT Assay
Resazurin Assay
LDH
2.2.2. BrdU Proliferation Assay
2.2.3. Analysis of Multicellular Aggregates (MCAs) Morphology in 3D Culture
Stereomicroscopy Analysis
Multicellular Aggregate Measurements
Morphology
MCA Immunocytochemical Analysis
MCA Electron Microscopy
3. Discussion
4. Materials and Methods
4.1. Cell Lines Cultivation
4.2. Chemicals and Cell Culture Reagents
4.3. Preussin (1)
4.4. Cell Exposures
4.4.1. Exposure in 2D Culture
4.4.2. Exposure in 3D Culture
4.5. Analysis of Cell Viability
4.5.1. 2D Culture
MTT Assay
4.5.2. 3D Culture
MTT Assay
Resazurin Reduction Assay
Lactate Dehydrogenase (LDH) Assay
4.6. Analysis of Cell Proliferation
4.6.1. 2D Culture
BrdU Assay
4.6.2. 3D Culture
4.7. Analysis of Cell Morphology in 3D Culture
4.7.1. MCA Measurements
4.7.2. Histological Analysis
4.7.3. Electron Microscopy
4.8. Immunocytochemistry in 3D Culture
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
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Malhão, F.; Ramos, A.A.; Buttachon, S.; Dethoup, T.; Kijjoa, A.; Rocha, E. Cytotoxic and Antiproliferative Effects of Preussin, a Hydroxypyrrolidine Derivative from the Marine Sponge-Associated Fungus Aspergillus candidus KUFA 0062, in a Panel of Breast Cancer Cell Lines and Using 2D and 3D Cultures. Mar. Drugs 2019, 17, 448. https://doi.org/10.3390/md17080448
Malhão F, Ramos AA, Buttachon S, Dethoup T, Kijjoa A, Rocha E. Cytotoxic and Antiproliferative Effects of Preussin, a Hydroxypyrrolidine Derivative from the Marine Sponge-Associated Fungus Aspergillus candidus KUFA 0062, in a Panel of Breast Cancer Cell Lines and Using 2D and 3D Cultures. Marine Drugs. 2019; 17(8):448. https://doi.org/10.3390/md17080448
Chicago/Turabian StyleMalhão, Fernanda, Alice A. Ramos, Suradet Buttachon, Tida Dethoup, Anake Kijjoa, and Eduardo Rocha. 2019. "Cytotoxic and Antiproliferative Effects of Preussin, a Hydroxypyrrolidine Derivative from the Marine Sponge-Associated Fungus Aspergillus candidus KUFA 0062, in a Panel of Breast Cancer Cell Lines and Using 2D and 3D Cultures" Marine Drugs 17, no. 8: 448. https://doi.org/10.3390/md17080448
APA StyleMalhão, F., Ramos, A. A., Buttachon, S., Dethoup, T., Kijjoa, A., & Rocha, E. (2019). Cytotoxic and Antiproliferative Effects of Preussin, a Hydroxypyrrolidine Derivative from the Marine Sponge-Associated Fungus Aspergillus candidus KUFA 0062, in a Panel of Breast Cancer Cell Lines and Using 2D and 3D Cultures. Marine Drugs, 17(8), 448. https://doi.org/10.3390/md17080448