Apoptpsis Of Melanoma Cell Induced By Essential Oil From Ginger Through MAPK Signaling Pathway And Its Mechanism

In the study,ginger essential oil extracted by supercritical CO₂extraction was added into mouse B16 melanoma cells at different concentrations and cultured.Determination of the apoptosis of mouse B16 melanoma cells under the influence of ginger essential oil,and the study of the related mechanism of ginger essential oil inducing the apoptosis of mouse B16 melanoma cells were experimented.The specific research contents are as follows:（1）The main components of ginger and ginger essential oil were identified and analyzed by gas chromatography-mass spectrometry,respectively.The results showed that the main volatile components in ginger wereα-curcumene,α-cedrene,γ-muurolene and camphene.The main components of ginger essential oil are zingiberene,6-gingerol,8-gingerol and gingerone.（2）The apoptosis of mouse B16 melanoma cells under the influence of ginger essential oil was determined by MTT test,cell morphology observation and flow cytometry analysis.The experimental results showed that ginger essential oil at a concentration of80-160μg/m L could affect the activity of mouse B16 melanoma cells.Mouse B16melanoma cells cultured for 24 h with various concentration of ginger essential oil showed shrinkage and rupture in different degrees.The results of cell cycle analysis showed that the cell cycle was blocked after 24 h treatment with ginger essential oil.The number of cells in G0/G1 phases decreased,while the number of cells in S and G2/M phase increased.This indicates that by adding ginger essential oil,the cell cycle of mouse B16 melanoma can be blocked in the S and G2/M phase,thus inhibiting the proliferation of cancer cells.（3）The apoptosis of mouse B16 melanoma cells cultured with ginger essential oil was detected by Hoechst33258 staining test,Annexin V-FITC/PI double staining test and mitochondrial membrane potential detection test.All the experimental results showed that mouse B16 melanoma cells could induce apoptosis of ginger essential oil,and the extent of apoptosis increased with the increase of ginger essential oil concentration.（4）The protein expression of Mitogen-activated protein kinase（MAPK）signaling pathway in the apoptosis of mouse B16 melanoma cells under the influence of ginger essential oil was determined by Western blotting assay.The results showed that 80,120 and160μg/m L ginger essential oil could induce the down-expression of extracellular regulated protein kinases（ERK）and p38 MAPK in mouse B16 melanoma cells,and increase c-Jun N-terminal kinase（JNK）expression.These results suggest that ginger essential oil may induce apoptosis of mouse B16 melanoma cells through MAPK signaling pathway,and reduce production of melanin.Besides,the expression of Bax and Bcl-2 proteins was measured.The results showed that the ratio of Bax to Bcl-2 proteins increased with the increase of concentration of ginger essential oil.This indicated that ginger essential oil can induce the apoptosis of mouse B16 cells.