Effect of S-Allyl –L-Cysteine on MCF-7 Cell Line 3-Mercaptopyruvate Sulfurtransferase/Sulfane Sulfur System, Viability and Apoptosis
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Chemicals
4.2. Cell Culture
4.3. Cell Treatment
4.4. Cytotoxicity of Reagents
4.5. Cell Viability by MTT Assay
4.6. Flow Cytometry (FC)
4.7. Western Blot Analysis
4.8. Densitometric Evaluation of Western Blot bands
4.9. Enzymes Assay
4.9.1. Cell Homogenization
4.9.2. MPST Activity
4.9.3. CTH Activity
4.9.4. Sulfane Sulfur
4.9.5. Protein
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AGE | Aged garlic extract |
ATCC | American Type Culture Collections |
casp-3 | caspase-3 |
casp-9 | caspase-9 |
CBS | cystathionine β-lyase |
CTH | γ-cystathionase |
CTL | control |
DTT | D,L-dithiothreitol |
NEM | N-ethylmaleimide |
FC | flow cytometry |
Gi | growth inhibition |
Hsp90 | heat shock protein 90 |
LDH | L-lactic dehydrogenase |
MGL | methionine γ-lyase |
MPST | 3-mercaptopyruvate sulfurtransferase |
MTT | 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide |
NADH | β-Nicotinamide adenine dinucleotide reduced disodium salt hydrate |
PA | pyruvate |
PCA | perchloric acid |
PLP | pyridoxal phosphate |
SAC | S-allyl- L-cysteine |
SAM | S-adenosyl-L-methionine |
SD | standard deviation |
WB | Western blot |
3MP | 3-mercaptopyruvate |
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24 h | 48 h | |
---|---|---|
% of Cytotoxicity | ||
800 µM | 0 | 0 |
1000 µM | 0 | 0 |
2245 µM | 0.2 | 0 |
MCF-7 Cell Line 24 h | MPST | CTH | Sulfane Sulfur |
---|---|---|---|
nmol·106·cells−1·min−1 | nmol·106·cells−1 | ||
CTL | 2714 ± 904 | 25 ± 9 | 826 ± 249 |
800 µM | 3042 ± 310 | 11 ± 6* | 949 ± 279 |
1000 µM | 3636 ± 818 | 5 ± 1* | 999 ± 198 |
2245 µM | 1828 ± 389 *Δ | 25 ± 16 | 497 ± 100 *Δ |
MCF-7 Cells 48 h | MPST | CTH | Sulfane Sulfur |
---|---|---|---|
nmol·106·cells−1·min−1 | nmol·106·cells−1 | ||
CTL | 3261 ± 798 | 30 ± 10 | 929 ± 194 |
800 µM | 2580 ± 761 | 50 ± 10* | 1731 ± 426* |
1000 µM | 4229 ± 830* | 10 ± 5* | 1905 ± 430* |
2245 µM | 2019 ± 382 Δ | 20 ± 1 | 571 ± 116*Δ |
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Bronowicka-Adamska, P.; Bentke, A.; Lasota, M.; Wróbel, M. Effect of S-Allyl –L-Cysteine on MCF-7 Cell Line 3-Mercaptopyruvate Sulfurtransferase/Sulfane Sulfur System, Viability and Apoptosis. Int. J. Mol. Sci. 2020, 21, 1090. https://doi.org/10.3390/ijms21031090
Bronowicka-Adamska P, Bentke A, Lasota M, Wróbel M. Effect of S-Allyl –L-Cysteine on MCF-7 Cell Line 3-Mercaptopyruvate Sulfurtransferase/Sulfane Sulfur System, Viability and Apoptosis. International Journal of Molecular Sciences. 2020; 21(3):1090. https://doi.org/10.3390/ijms21031090
Chicago/Turabian StyleBronowicka-Adamska, Patrycja, Anna Bentke, Małgorzata Lasota, and Maria Wróbel. 2020. "Effect of S-Allyl –L-Cysteine on MCF-7 Cell Line 3-Mercaptopyruvate Sulfurtransferase/Sulfane Sulfur System, Viability and Apoptosis" International Journal of Molecular Sciences 21, no. 3: 1090. https://doi.org/10.3390/ijms21031090