Triterpene Acid (3-O-p-Coumaroyltormentic Acid) Isolated From Aronia Extracts Inhibits Breast Cancer Stem Cell Formation through Downregulation of c-Myc Protein
Abstract
:1. Introduction
2. Results
2.1. Isolation of the CSC Inhibitor From Aronia
2.2. Structure Determination of the Isolated CSC Inhibitor
2.3. 3-O-p-Coumaroyltormentic Acid Inhibits Breast Cancer Cell Proliferation and Mammosphere Formation
2.4. Treatment with 3-O-trans-p-Coumaroyltormentic Acid Decreased the CD44high/CD24low-Expressing Subpopulation and the Proportion of ALDH-Positive Breast Cancer Cells
2.5. Effect of 3-O-Trans-p-Coumaroyltormentic Acid on the Protein Expression Levels of c-Myc in Breast CSCs
2.6. c-Myc is a Survival Factor for Breast CSC Formation
2.7. 3-O-trans-p-Coumaroyltormentic Acid Inhibits the Expression of Self-Renewal Genes in CSCs and the Proliferation of Mammospheres
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Plant Material
4.3. Extraction and Isolation
4.4. Structure Analysis of Purified Sample
4.5. Culture of Human Breast Cancer Cells and the Formation of Mammospheres
4.6. Automated Counting of Mammospheres
4.7. Cell Proliferation Assay
4.8. Flow Cytometric Analysis of CD44 and CD24 Expression
4.9. Clonogenic Assay
4.10. Scratch Migration Assay
4.11. Real-Time RT-PCR
4.12. ALDEFLUOR Assay
4.13. Western Blotting
4.14. Small Interfering RNA (siRNA)
4.15. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
Abbreviations
CSCs | Cancer stem cells |
AMJ | Aronia melanocarpa juice |
NICE | NIST’s integrated colony enumerator |
ESI | Electrospray ionization |
NMR | Nuclear magnetic resonance |
COSY | Correlation spectroscopy |
HMQC | Heteronuclear multiple-quantum correlation spectroscopy |
HMBC | Heteronuclear multiple-bond correlation spectroscopy |
ROI | Regions of interest |
MFE | Mammosphere formation efficiency |
Trans | trans-Coumaroyltormentic acid |
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Choi, H.S.; Kim, S.-L.; Kim, J.-H.; Deng, H.-Y.; Yun, B.-S.; Lee, D.-S. Triterpene Acid (3-O-p-Coumaroyltormentic Acid) Isolated From Aronia Extracts Inhibits Breast Cancer Stem Cell Formation through Downregulation of c-Myc Protein. Int. J. Mol. Sci. 2018, 19, 2528. https://doi.org/10.3390/ijms19092528
Choi HS, Kim S-L, Kim J-H, Deng H-Y, Yun B-S, Lee D-S. Triterpene Acid (3-O-p-Coumaroyltormentic Acid) Isolated From Aronia Extracts Inhibits Breast Cancer Stem Cell Formation through Downregulation of c-Myc Protein. International Journal of Molecular Sciences. 2018; 19(9):2528. https://doi.org/10.3390/ijms19092528
Chicago/Turabian StyleChoi, Hack Sun, Su-Lim Kim, Ji-Hyang Kim, Hong-Yuan Deng, Bong-Sik Yun, and Dong-Sun Lee. 2018. "Triterpene Acid (3-O-p-Coumaroyltormentic Acid) Isolated From Aronia Extracts Inhibits Breast Cancer Stem Cell Formation through Downregulation of c-Myc Protein" International Journal of Molecular Sciences 19, no. 9: 2528. https://doi.org/10.3390/ijms19092528