Cytotoxic and Pro-Apoptotic Effects of Cassane Diterpenoids from the Seeds of Caesalpinia sappan in Cancer Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. Identification of New Compounds
2.2. Cytotoxicity Assay
2.3. Compound 1 Mediates G1 Cell Cycle Arrest
2.4. Pro-apoptotic and p53 Suppressing Activities of Compound 1
3. Materials and Methods
3.1. General Procedures
3.2. Plant Material
3.3. Extraction and Isolation
3.4. Spectroscopic Data
3.5. Reagents
3.6. Cell Culture
3.7. MTT Assay
3.8. Cell Cycle Analysis
3.9. Hoechst 33342 Staining Assay
3.10. Annexin V/PI Staining Assay
3.11. Western Blot Analysis
3.12. Statistical Analysis
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
CD | Circular Dichroism |
13C-NMR | 13C Nuclear Magnetic Resonance |
1D-NMR | 1 Dimension Nuclear Magnetic Resonance |
2D-NMR | 2 Dimension Nuclear Magnetic Resonance |
DMSO | Dimethyl Sulphoxide |
ESIMS | Electrospray Ionization Mass Spectrometry |
FBS | Fetal Bovine Serum |
HMBC | Heteronuclear Multiple Bond Correlation |
1H-NMR | 1H Nuclear Magnetic Resonance |
HRESIMS | High-resolution ESIMS |
HSQC | Heteronuclear Single-quantum Correlation |
IR | Infrared Spectroscopy |
MTT | 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium Bromide |
PBS | Phosphate-buffered Saline |
Preparative HPLC | Preparative High Performance Liquid Chromatography |
ROESY | Rotating-frame Nuclear Overhauser Enhancement Spectroscopy |
SDS | Sodium Dodecyl Sulfate |
TLC | Thin Layer Chromatography |
UV | Ultraviolet Spectroscopy |
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- Sample Availability: Samples of the compounds 1–12 are available from the authors.
Position | 1 | 2 | 3 | 4 | 5 |
---|---|---|---|---|---|
1α | 1.13, m | 1.14, dd (13.1, 4.8) | 1.48, m | 1.20, m | 1.26, m |
1β | 2.27, m | 1.75, m | 1.78, m | 2.05, m | 1.46, m |
2α | 1.60, m | 1.87, m | 1.61, m | 1.64, m | 1.62, m |
2β | 1.44, m | 1.68, m | 2.01, m | 2.28, m | 2.31, m |
3α | 1.77, m | 1.63, m | 1.56, m | 1.93, m | 1.94, m |
3β | 1.61, m | 2.42, dd (13.0, 1.3) | 2.01, m | 2.05, m | 2.00, m |
5 | 1.81, m | 2.01, dd (12.1, 2.3) | 1.83, m | 1.68, m | 1.65, m |
6α | 1.21, m | 1.45, m | 1.57, m | 1.21, m | 1.24, m |
6β | 1.41, m | 1.64, m | 1.75, m | 2.10, m | 1.62, m |
7α | 1.40, m | 1.57, m | 1.19, m | 1.36, m | 1.36, m |
7β | 1.70, m | 1.76, m | 1.76, m | 1.56, m | 1.67, m |
8 | 1.88, m | 1.78, m | 2.17, m | 2.32, m | 2.34, m |
9 | 1.61, m | 1.56, m | 1.83, m | 1.50, m | 2.24, m |
11α | 2.71, dd (16.8, 6.4) | 2.60, dd (16.9, 6.8) | 4.73, d (3.1) | 2.60, m | 2.25, m |
11β | 2.34, m | 2.35, dd (16.9, 11.8) | - | 1.74, m | 2.36, m |
12 | - | - | - | 4.83, dd (11.6, 6.0) | - |
14 | 2.61, m | 2.62, m | 2.66, m | 2.92, m | 2.38, m |
15 | 6.18, d (1.7) | 6.18, d (1.7) | 6.23, d (1.7) | 5.67, s | α: 3.18, d (18.6) β: 3.01, d (18.6) |
16 | 7.21, d (1.7) | 7.22, d (1.7) | 7.32, d (1.7) | - | - |
17 | 0.97, d (7.0) | 0.99, d (7.1) | 0.98, d (7.1) | 1.04, d (7.1) | 0.88, d (7.0) |
19α | 1.29, s | - | 3.50, d (12.5) | 3.69, d (12.0) | 3.72, d (11.7) |
19β | 4.89, d (12.6) | 4.33, dd (11.9, 2.1) | 4.35, dd (11.8, 2.5) | ||
20 | α: 3.86, d (12.1) β: 3.97, d (12.1) | 0.74, s | 4.98, s | 4.83, d (2.1) | 4.98, s |
18-OMe | 3.68, s | 3.71, s | 3.72, s | 3.67, s | 3.67, s |
19-OMe | - | 3.74, s | - | - | - |
Position | 1 | 2 | 3 | 4 | 5 |
---|---|---|---|---|---|
1 | 31.4 | 39.1 | 34.9 | 37.8 | 37.8 |
2 | 17.6 | 19.1 | 17.9 | 21.0 | 20.9 |
3 | 35.4 | 34.7 | 35.7 | 35.7 | 35.6 |
4 | 49.4 | 57.6 | 47.3 | 45.6 | 45.6 |
5 | 51.5 | 50.5 | 46.2 | 45.1 | 45.1 |
6 | 23.5 | 25.7 | 27.8 | 23.9 | 23.5 |
7 | 30.5 | 31.7 | 28.1 | 29.0 | 21.9 |
8 | 36.6 | 36.1 | 38.1 | 41.2 | 36.5 |
9 | 45.0 | 45.2 | 45.1 | 41.7 | 41.9 |
10 | 40.6 | 37.0 | 43.4 | 38.7 | 38.2 |
11 | 23.1 | 22.5 | 70.1 | 33.8 | 29.2 |
12 | 149.6 | 149.5 | 146.7 | 79.6 | 149.1 |
13 | 122.1 | 122.5 | 129.5 | 175.5 | 115.6 |
14 | 31.4 | 31.7 | 32.6 | 36.9 | 32.2 |
15 | 109.7 | 109.7 | 109.5 | 110.9 | 34.9 |
16 | 140.3 | 140.6 | 143.1 | 174.0 | 176.9 |
17 | 16.9 | 17.8 | 14.2 | 13.3 | 14.3 |
18 | 179.2 | 173.8 | 176.1 | 177.2 | 175.7 |
19 | 19.1 | 172.6 | 67.1 | 61.5 | 61.9 |
20 | 61.2 | 13.7 | 106.2 | 96.8 | 97.3 |
18-OMe | 52.0 | 52.8 | 52.2 | 51.8 | 51.8 |
19-OMe | 52.0 |
Compounds | A2780 | HEY | AGS | A549 |
---|---|---|---|---|
1 | 10.4% ± 4.7% | 10.2% ± 9.8% | 4.9% ± 1.3% | 32.9% ± 13.0% |
2 | 92.0% ± 7.1% | 95.7% ± 6.4% | 80.8% ± 9.7% | 80.0% ± 8.0% |
3 | 74.8% ± 8.2% | 79.9% ± 12.7% | 71.7% ± 18.2% | 74.6% ± 5.4% |
4 | 97.3% ± 7.4% | 95.2% ± 6.0% | 80.8% ± 15.7% | 79.9% ± 10.0% |
5 | 91.7% ± 1.4% | 97.2% ± 2.8% | 83.0% ± 16.6% | 84.9% ± 10.0% |
6 | 95.2% ± 1.0% | 91.3% ± 9.6% | 82.3% ± 9.9% | 83.5% ± 7.6% |
7 | 37.9% ± 5.6% | 68.2% ± 5.5% | 69.5% ± 8.9% | 73.5% ± 12.0% |
8 | 49.5% ± 5.8% | 41.6% ± 9.0% | 14.6% ± 2.3% | 50.9% ± 12.3% |
9 | 64.4% ± 4.6% | 74.1% ± 7.0% | 71.1% ± 12.7% | 72.8% ± 4.7% |
10 | 57.4% ± 4.5% | 71.4% ± 1.9% | 62.7% ± 13.5% | 75.2% ± 9.8% |
11 | 95.4% ± 2.7% | 91.7% ± 6.1% | 85.8% ± 9.4% | 74.8% ± 13.8% |
12 | 92.1% ± 6.6% | 97.5% ± 2.0% | 82.1% ± 14.0% | 82.2% ± 10.6% |
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Share and Cite
Bao, H.; Zhang, L.-L.; Liu, Q.-Y.; Feng, L.; Ye, Y.; Lu, J.-J.; Lin, L.-G. Cytotoxic and Pro-Apoptotic Effects of Cassane Diterpenoids from the Seeds of Caesalpinia sappan in Cancer Cells. Molecules 2016, 21, 791. https://doi.org/10.3390/molecules21060791
Bao H, Zhang L-L, Liu Q-Y, Feng L, Ye Y, Lu J-J, Lin L-G. Cytotoxic and Pro-Apoptotic Effects of Cassane Diterpenoids from the Seeds of Caesalpinia sappan in Cancer Cells. Molecules. 2016; 21(6):791. https://doi.org/10.3390/molecules21060791
Chicago/Turabian StyleBao, Han, Le-Le Zhang, Qian-Yu Liu, Lu Feng, Yang Ye, Jin-Jian Lu, and Li-Gen Lin. 2016. "Cytotoxic and Pro-Apoptotic Effects of Cassane Diterpenoids from the Seeds of Caesalpinia sappan in Cancer Cells" Molecules 21, no. 6: 791. https://doi.org/10.3390/molecules21060791