| Objectives This study aims to investigate the anti-tumor effect of ?-Mangostin on melanoma cells, evaluate the synergistic therapeutic effect of ?-Mangostin in combination with chemotherapeutics on melanoma cells, and probe the possiblely involved and related signaling pathways.Methods 1. MTT assay was performed to investigate the proliferation inhibition of ?-Mangostin on melanoma cells; 2. Transmision electron microscopy was performed to observe the morphological changes of melanoma cells treated with ?-Mangostin, and JC-1 staining was used to detect its effect on mitochondrial membrane potential; 3. Flow cytometry analysis of apoptosis and cell cycle were utilized to detect the effect of ?-Mangostin on cell apoptosis induction and cell cycle arrest, respectively; 4. Migration assay and invasion assay were performed to detect its influence on the cell migration and invasion ability, respectively; 5. The synergistic anti-melanoma effect with common chemotherapy drugs in vitro were detected by MTT method; 6. Western Blot and Real time-PCR were carried out to analyze the influence of ?-Mangostin on some of the common signaling pathways in melanoma cell lines; 7. C57BL/6 mice subcutaneous transplantation tumor model was built to evaluate the anti-tumor effect of ?-Mangostin against melanoma, and the effect of ?-Mangostin combined with common chemotherapy drugs were detected in vitro.The signaling ways were further tested in the dissected tumor tissues.Results: 1. ?-Mangostin can significantly reduce the proliferation rate in humanmelanoma cell line A375, M14, SK- MEL- 2 and mice melanoma cell line B16F10; 2. Upon ?-Mangostin treatment, electron microscope analysis showed Mitochondrial mitochondrial membrane structure destruction and mitochondrial swelling in various melanoma cells lines, as well as the increasing ratios of F540/F590 when treated with 5 and 10 ?M ?-Mangostin(1.63 and 3.00?4.38 and 20.25?3.73 and 6.9?8.67 and 20.6 folds, respectively, p<0.05); 3. 10 and 20 ?M ?-Mangostin can increase the apoptosis rate of melanoma cells by 7.89% and 70.1%, 29.57% and 85.92%, 19.52% and 86.49%, 19.67% and 47.1% respectively(p < 0.05), and can make the melanoma cell cycle arrest before G1 phase;4. The number of cells migrated were significantely reduced when treated with 5 and 10 ?M(0.48 and 0.24, 0.48 and 0.24, 0.72 and 0.44, 0.49 and 0.16 folds respectively, p < 0.01), as well as the number of cells invaded into the lower room(0.35 and 0.32, 0.37 and 0.24, 0.37 and 0.31, 0.47 and 0.16 folds, p < 0.01); 5. ?-Mangostin can synergy with cisplatin and dacarbazine respectively in melanoma cells; 6. ?-Mangostin can inhibit the level of RAS protein and m RNA expression, and the level of phosphorylation of PI3 K, as well as the level of MITF protein and m RNA expression in M14 cells. 7. ?-Mangostin treatment can inhibit tumor growth significantly. The 10 th day tumor size of ?-Mangostin group is such smaller than that of the control group(955.10±103.10 versus 336.10±35.95 mm3, p < 0.001). The tumors of alpha Mangostin and cisplatin combination group are smaller than that of ?-Mangostin group and cisplatin group. The sizes are 164.80±23.02, 336.00±35.95 and 583.00±83.32 cm3 respectively(p < 0.01). ?-Mangostin group shows the lower level of the Ras protein expression and the PI3 K phosphorylation compared with the control group.Conclusion: ?-Mangostin has antitumor effect on melanoma cells both in vivo and in vitro. The underlying mechanism may involve the inhibition of Ras, MITF and PI3 K. ?-Mangostin can synergize the inhibitory effect of cisplatin and dacarbazine on melanoma cells. |
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