Effect Of Turmeric Volatile Oil On Proliferation And Apoptosis Of Melanoma Wm9 Cells

Objective: To investigate the effect of turmeric volatile oil on the proliferation of melanoma wm9 cells,and explore the mechanism of its apoptosis。 Methods: Different concentration of turmeric volatile oil were applied on melanoma wm9 cells;Detection of proliferation inhibition rate by CCK-8(Cell Counting Kit-8);Morphological changes of cells were observed by inverted microscope;Caspase-3 activities were evaluated by enzyme linked immunosorbent assay;Cell cycle and apoptosis was analyzed with flow cytometry(FCM)。Results: To investigate the activities of turmeric volatile oil with different concentrations(0, 5, 10, 20, 40 umol / L) on human melanoma wm9 cells after cultured for 24 h, 48 h, 72 h By cell counting kits(CCK-8). It showed increasing inhibitory effect with drug concentration ranged from low to high at time-dose dependent manner. after treated for 48 h, it showed the most obvious inhibition effect When drug concentration reached at 40mg/L. Irregular cell morphology were presented under inverted microscope after treated with turmeric volatile oil, and the cell proliferation speed of wm9 cells were declined as well. When the drug concentration was up to 20 mg / L, there was karyopyknosis, part of the tumor cell membrane began to break down and part of the contents of cells were released. When the drug concentration reached at 40 mg / L, the normal structure of the tumor cells was unable to see, and large amounts of cellular debris can be seen as well; Caspase-3 enzyme activity assay showed that turmeric volatile oil can induce the apoptosis of melanoma wm9, it showed that positive correlation between drug concentration and Caspase-3 enzyme activity with the increasing of drug concentration. When the drug concentration reached at 40mg/L, the caspase-3 enzyme activity reached at the maximum. the capability of apoptosis-inducing on melanoma wm9 cells were different after treated with turmeric volatile oil for 48 hours. With the increasing of drug concentration,there was positive correlation between drug concentration and apoptosis rate, early apoptosis rates were(12.30 + 0.01),(11.95 + 0.04),(14.63 + 0.12), and 25.74 + 0.20%, respectively, while the late apoptosis rates were(10.18 + 0.02),(13.74 + 0.13),(35.68 + 0.21), and(58.14 + 0.13%)respectively, which we can see the early and late apoptosis rate reached the maximum value when drug concentration at 40mg/L, and in the same drug concentration, late apoptosis rate is higher than early apoptosis rate, it has statistically significant differences compared with the control group(P < 0.05). Cell cycle detection showed that G1, S phase cells gradually increased with the increasing of drug concentration, while the number of G2 cells gradually decreased. It showed negative correlation between drug and cell numbers. With the increasing of drug concentration, the number of G2 phase cells gradually increased, which indicated that the cells were blocked in G2 / M phase. Conclusion: turmeric volatile oil has an effect of proliferation inhibition and apoptosis-inducing on melanoma wm9 cells. The mechanism of apoptosis may be related to Caspase-3 enzyme, which activates cell apoptosis, moreover, normal cell cycle was blocked, thereby the proliferation and differentiation of tumor cells were inhibited.