Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line
Abstract
:1. Introduction
2. Results
2.1. The Effect of Cell Density on Cell Viability
2.2. SD Reduces Viability of the B16F10 Melanoma Cells
2.3. SD Inhibits de Novo DNA Synthesis
2.4. SD Enhances DNA Fragmentation
2.5. Effect of SD on the Total Protein Contents in Melanoma Cells
2.6. SD Inhibits the Levels of Protein Markers for Cell Proliferation
Sarcophine-diol concentration (µM) | |||||||||
---|---|---|---|---|---|---|---|---|---|
0 | 62.5 | 125 | 250 | 0 | 62.5 | 125 | 250 | ||
24 h treatment | 72 h treatment | ||||||||
Proliferation markers | STAT-3 | 100% | 78.8 ± 9.5 n = 4 | 62.9 ± 10.4 n = 4 (*) | 36.0 ± 6.0 n = 4 (*) | 100% | 70.4 ± 18.2 n = 3 | 59.8 ± 8.4 n = 3 (*) | 41.8 ± 10.5 n = 3 (*) |
Cdk4 | 100% | 105.3 ± 9.2 n = 3 | 92.8 ± 14.3 n = 3 | 101.8 ± 15.3 n = 3 | 100% | 112.5 ± 27.2 n = 3 | 97.4 ± 18.9 n = 3 | 118.7 ± 20.1 n = 3 | |
Cyclin-D1 | 100% | 75.3 ± 10.5 n = 3 | 35.5 ± 7.2 n = 3 (*) | 7.7 ± 14.2 n = 3 (*) | 100% | 87.8 ± 12.6 n = 3 | 48.9 ± 9.8 n = 3 (*) | 4.9 ± 6.2 n = 3 (*) | |
Cyclin-D2 | ND | ND | ND | ND | ND | ND | ND | ND | |
Apoptosis markers | C-PARP | 100% | 123.2 ± 18.5 n = 3 | 132.1 ± 18.1 n = 3 (*) | 168.3 ± 24.1 n = 3 (*) | 100% | 189.3 ± 32.2 n = 3 (*) | 216.0 ± 42.7 n = 3 (*) | 276.0 ± 56.8 n = 3 (*) |
p53 | 100% | 128.0 ± 23.9 n = 3 | 132.1 ± 19.6 n = 3 (*) | 132.7 ± 16.9 n = 3 (*) | 100% | 141.5 ± 33.3 n = 3 | 159.9 ± 22.6 n = 3 (*) | 177.6 ± 19.4 n = 3 (*) | |
C-Casp-9 | 100% | 124.3 ± 27.7 n = 3 | 132.3 ± 20.5 n = 3 (*) | 147.4 ± 18.1 n = 3 (*) | 100% | 130.6 ± 23.5 n = 3 | 158.0 ± 31.6 n = 3 (*) | 179.1 ± 18.9 n = 3 (*) | |
C-Casp-3 | 100% | 120.2 ± 14.3 n = 3 | 133.3 ± 9.9 n = 3 (*) | 140.3 ± 16.7 n = 3 (*) | 100% | 128.2 ± 25.5 n = 3 | 141.7 ± 22.5 n = 3 (*) | 174.2 ± 21.0 n = 3 (*) | |
C-Casp-8 | 100% | 119.5 ± 22.6 n = 3 | 132.3 ± 25.3 n = 3 (*) | 160.3 ± 18.7 n = 3 (*) | 100% | 123.9 ± 25.1 n = 3 | 178.3 ± 33.6 n = 3 (*) | 213 ± 51.2 n = 3 (*) |
2.7. SD Increases the Expression Levels of Protein Markers for Apoptosis
2.8. SD Increases Enzymatic Activities of the Cleaved-Caspases-3, -8 and -9
2.9. Discussion
3. Experimental Section
3.1. Materials and Reagents
3.2. Synthesis of SD
3.3. Cell Culture
3.4. Total Cellular Protein Extract
3.5. Determination of Protein Content
3.6. Western Blot Analyses of Protein Markers for Cell Proliferation and Apoptosis
3.7. Cell Viability Assays
3.7.1. Staining of the Cells with Trypan-Blue
3.7.2. MTT Assay
3.8. Determination of DNA Synthesis
3.9. Determination of DNA Fragmentation in Situ
3.10. Determination of Enzymatic Activities of the Cleaved-Caspases-3, -8 and -9
3.11. Statistical Analysis
Acknowledgements
- Samples Availability: Available from the authors.
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Szymanski, P.T.; Kuppast, B.; Ahmed, S.A.; Khalifa, S.; Fahmy, H. Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line. Mar. Drugs 2012, 10, 1-19. https://doi.org/10.3390/md10010001
Szymanski PT, Kuppast B, Ahmed SA, Khalifa S, Fahmy H. Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line. Marine Drugs. 2012; 10(1):1-19. https://doi.org/10.3390/md10010001
Chicago/Turabian StyleSzymanski, Pawel T., Bhimanna Kuppast, Safwat A. Ahmed, Sherief Khalifa, and Hesham Fahmy. 2012. "Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line" Marine Drugs 10, no. 1: 1-19. https://doi.org/10.3390/md10010001