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, 2014, 378684

Role of JNK Activation and Mitochondrial Bax Translocation in Allicin-Induced Apoptosis in Human Ovarian Cancer SKOV3 Cells


Role of JNK Activation and Mitochondrial Bax Translocation in Allicin-Induced Apoptosis in Human Ovarian Cancer SKOV3 Cells

Ling Xu et al. Evid Based Complement Alternat Med.


Background. Allicin, the major component of freshly crushed garlic, is one of the most biologically active compounds of garlic; it has been reported to induce apoptosis in cancer cells; however, the mechanism by which allicin exerts its apoptotic effects is not fully understood. The aim of the present study was to further elucidate the apoptotic pathways induced by allicin in the human ovarian cancer cell line SKOV3. Methods. Cell proliferation and apoptosis were measured by cell-counting assay and flow cytometry analysis. Activation of the signaling pathway was screened by human phospho-kinase array analysis, and the activated pathway and its related proteins were further confirmed by western blot analysis. Results. Allicin induced SKOV3 cell apoptosis and JNK phosphorylation in a time- and dose-dependent manner, but these were significantly blocked by SP600125 (an inhibitor of JNK). The findings suggest that JNK phosphorylation is related to the action of allicin on SKOV3 cells. Furthermore, JNK activation induced Bcl-2 family activation, triggered mitochondria-mediated signaling pathways, and led to the translocation of a considerable amount of Bax and cytochrome c release. Conclusions. JNK activation and mitochondrial Bax translocation are involved in allicin-induced apoptosis in SKOV3 cells. Our data input new insights to the literature of allicin-induced apoptosis.


Figure 1
Figure 1
Inhibitory effect of allicin on SKOV3 cell proliferation. SKOV3 cells were treated with various doses of allicin for 24, 48, and 72 h. Cell proliferation was determined using cell-counting assay and expressed as the percentage of the absorbance value obtained without allicin.
Figure 2
Figure 2
Flow cytometry analysis of allicin and/or SP600125 in SKOV3 cell apoptosis. SKOV3 cells were pretreated with 20 μM SP600125 for 30 min before incubation with 25 μg/mL of allicin, and apoptotic cells were measured by cytometry after 48 h. Data (mean ± SD) are representative of three experiments. (a) is a representative figure and (b) is a statistical graph. Asterisks indicate statistically significant difference (**P < 0.01).
Figure 3
Figure 3
Effect of allicin and/or SP600125 on the phosphorylation of JNK in SKOV3 cells. (a) Treatment with various concentrations of allicin for 15 min. (b) Treatment with 25 μg/mL of allicin at indicated times. (c) Pretreatment with 20 μM SP600125 for 30 min before incubation with 25 μg/mL of allicin for 15 min; JNK phosphorylation was measured by western blot analysis after 48 h.
Figure 4
Figure 4
Western blot analysis showing cytochrome c and Bax levels in response to allicin. (a) SKOV3 cells were treated with 25 μg/mL of allicin for 12 h. Subsequently, cytosolic and mitochondrial fractions were prepared and western blot analysis was carried out (20 μg of protein) as described in Materials and Methods. (b) Pretreatment with or without the JNK inhibitor SP600125 for 30 min, followed by treatment with allicin for 12 h to analyze Bax and cytochrome c. Data are representative of three independent experiments showing a similar pattern of expression. β-Actin and Hps60 were used as internal control.

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