A New Highlight of Ephedra alata Decne Properties as Potential Adjuvant in Combination with Cisplatin to Induce Cell Death of 4T1 Breast Cancer Cells In Vitro and In Vivo
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Lines
2.2. Drugs, Antibodies, and Chemical Reagents
2.3. Preparation of Ephedra alata Extract
2.4. LC–MS/MS Analysis
2.5. Cell Viability Assays
2.6. Combination Index Analysis
2.7. Apoptosis Analysis
2.8. Flow Cytometry Analysis of Apoptosis
2.9. Western Blotting
2.10. Preparation of Mitochondrial and Cytosolic Protein Extracts
2.11. Mouse Strain and Tumor Growth Experiments
2.12. Biochemical Assays and Analysis
2.13. Statistical Analysis
3. Results
3.1. Identification and Quantification of Phenolic Compounds in E.A. Extracts
3.2. E.A. Extract Decreased Breast Cancer Cells Viability
3.3. E.A. Synergized with CDDP to Reduce 4T1 Breast Cancer Cell Viability
3.4. E.A. Synergized with CDDP to Enhance Apoptosis in 4T1 Breast Cancer Cells
3.5. E.A. Sensitized Breast Cancer Cells to CDDP-Mediated Caspase Activation
3.6. E.A./CDDP Combination-Induced Apoptosis through a Modulation of Bcl-2 Proteins Family
3.7. E.A./CDDP Combination Induced Smac/Diablo and Cytochrome C Release from the Mitochondria
3.8. E.A./CDDP Combination Synergized to Prevent Murine Mammary Carcinoma Growth
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Compound | Content (%) |
---|---|
Caffeic acid derivative | 0.54 |
Citric acid | 0.28 |
Syringic acid hexoside | 0.70 |
Syringic acid | 0.73 |
Gallocatechin | 0.40 |
Ferulic acid | 0.19 |
Quercetin dihexoside | 1.02 |
Vicenin-2 | 9.33 |
Quercetin-3-O-galactoside | 1.95 |
Isoorientin-4-O-glucoside | 1.74 |
6-Hydroxyquercetin-3-O-di-hexose | 14.67 |
Quercetin 3-O-glucoside | 2.48 |
Luteolin-8-C-β-d-glucopyranoside | 0.18 |
Quercetin | 19.85 |
Myricetin hexoside | 8.59 |
Quercetin 3-O-[6”-(3-hydroxyl-3-methylglutaryl)-β-d-galactoside] | 24.48 |
Myricetin-3-O-glucoside | 3.05 |
Quercetin 3-O-rhamnoside | 0.76 |
7-O-rhamnoside | |
Kaempferol-O-di-deoxyhexoside | 1.08 |
IC50 | 24 h | 48 h | 72 h |
---|---|---|---|
E.A. | - | 65 µg/mL | 30 µg/mL |
CDDP | 20 µM | 10 µM | 3 µM |
Ratio Cytosol/Mitochondria Fractions | |||
---|---|---|---|
Bax | Smac/Diablo | Cytochrome c | |
E.A. | 2.1 | 0.38 | 0.91 |
CCDP 5 µM | 0.62 | 0.43 | 0.45 |
CCDP 10 µM | 0.25 | 0.90 | 0.55 |
E.A. + CCDP 5 µM | 0.12 | 5.10 | 0.41 |
E.A. + CCDP 10 µM | 0.10 | 9.69 | 0.66 |
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Sioud, F.; Amor, S.; Toumia, I.b.; Lahmar, A.; Aires, V.; Chekir-Ghedira, L.; Delmas, D. A New Highlight of Ephedra alata Decne Properties as Potential Adjuvant in Combination with Cisplatin to Induce Cell Death of 4T1 Breast Cancer Cells In Vitro and In Vivo. Cells 2020, 9, 362. https://doi.org/10.3390/cells9020362
Sioud F, Amor S, Toumia Ib, Lahmar A, Aires V, Chekir-Ghedira L, Delmas D. A New Highlight of Ephedra alata Decne Properties as Potential Adjuvant in Combination with Cisplatin to Induce Cell Death of 4T1 Breast Cancer Cells In Vitro and In Vivo. Cells. 2020; 9(2):362. https://doi.org/10.3390/cells9020362
Chicago/Turabian StyleSioud, Fairouz, Souheila Amor, Imène ben Toumia, Aida Lahmar, Virginie Aires, Leila Chekir-Ghedira, and Dominique Delmas. 2020. "A New Highlight of Ephedra alata Decne Properties as Potential Adjuvant in Combination with Cisplatin to Induce Cell Death of 4T1 Breast Cancer Cells In Vitro and In Vivo" Cells 9, no. 2: 362. https://doi.org/10.3390/cells9020362