Sarcophine-Diol Inhibits Expression of COX-2, Inhibits Activity of cPLA2, Enhances Degradation of PLA2 and PLCγ1 and Inhibits Cell Membrane Permeability in Mouse Melanoma B16F10 Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. Wound Healing Assay
2.2. SD Inhibits Cell Multiplication
Time 0 | 0 µM | 62.5 µM | 125 µM | 250 µM | |
---|---|---|---|---|---|
Total protein content (µg) | 26.62 ± 5.49, n = 5 | 309.41 ± 43.93, n = 5 | 201.67 ± 35.35, n = 5 * | 165.83 ± 19.57, n = 5 * | 38.57 ± 7.32, n = 5 * |
Total living cells | 1.86 × 105 ± 4.7 × 104, n = 5 | 2.4 × 106 ± 4.5 ×105, n = 5 | 1.68 × 106 ± 2.7 × 105, n = 5 * | 1.11 × 106 ± 1.9 × 105, n = 5 * | 2.3 × 105 ± 5.5 × 104, n = 5 * |
2.3. SD Inhibits Cell Membrane Permeability
2.4. SD Inhibits Expression Levels of Cox-2 and Enhances Degradation of PLA2 and PLCγ1
Control | 24 h, SD | 72 h, SD | |
---|---|---|---|
Cox-2 | 100 | 56.81 ± 10.22, n = 4, * | 27.62 ± 6.85, n = 4, * |
PLA2 | 100 | 57.41 ± 14.63, n = 7, * | 30.58 ± 9.67, n = 7, * |
PLCγ1 | 100 | 59.20 ± 6.53, n = 8, * | 34.44 ± 10.05, n = 8, * |
i-PARP | 100 | 68.51 ± 12.06, n = 4, * | 50.16 ± 13.47, n = 4, * |
c-PARP | 100 | 150.41 ± 26.31, n = 4, * | 226.32 ± 35.27, n = 4, * |
c-Casp-3 | 100 | 143.1 ± 20.26, n = 6, * | 197.22 ± 21.75, n = 6, * |
2.5. SD Inhibits Enzymatic Activity of cPLA2
2.6. Discussion
3. Experimental Section
3.1. Chemical and Reagents
3.2. Semi-Synthesis of Sarcophine-Diol (SD)
3.3. Cell Culture
3.4. Wound Healing Assay
3.5. Cell Membrane Permeability
3.6. Determination of Cell Multiplication and Cellular Protein Content
3.7. Western Blot Analysis
3.8. Determination of Enzymatic Activity of cPLA2
3.9. Staining of the Cells with Trypan-Blue
3.10. Determination of Protein Content
3.11. Statistical Analysis
4. Conclusions
Acknowledgments
References
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Szymanski, P.T.; Muley, P.; Ahmed, S.A.; Khalifa, S.; Fahmy, H. Sarcophine-Diol Inhibits Expression of COX-2, Inhibits Activity of cPLA2, Enhances Degradation of PLA2 and PLCγ1 and Inhibits Cell Membrane Permeability in Mouse Melanoma B16F10 Cells. Mar. Drugs 2012, 10, 2166-2180. https://doi.org/10.3390/md10102166
Szymanski PT, Muley P, Ahmed SA, Khalifa S, Fahmy H. Sarcophine-Diol Inhibits Expression of COX-2, Inhibits Activity of cPLA2, Enhances Degradation of PLA2 and PLCγ1 and Inhibits Cell Membrane Permeability in Mouse Melanoma B16F10 Cells. Marine Drugs. 2012; 10(10):2166-2180. https://doi.org/10.3390/md10102166
Chicago/Turabian StyleSzymanski, Pawel T., Pratik Muley, Safwat A. Ahmed, Sherief Khalifa, and Hesham Fahmy. 2012. "Sarcophine-Diol Inhibits Expression of COX-2, Inhibits Activity of cPLA2, Enhances Degradation of PLA2 and PLCγ1 and Inhibits Cell Membrane Permeability in Mouse Melanoma B16F10 Cells" Marine Drugs 10, no. 10: 2166-2180. https://doi.org/10.3390/md10102166