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

Xu L, Yu J, Zhai D, Zhang D, Shen W, Bai L, Cai Z, Yu C - Evid Based Complement Alternat Med (2014)

Bottom Line: Results.Conclusions.Our data input new insights to the literature of allicin-induced apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Traditional Chinese Gynecology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus

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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fig4: 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.

Mentions: The Bax (2D2) and cytochrome c levels in the mitochondrial and cytosolic fractions were examined to further elucidate whether the JNK pathway is involved in downstream molecular events of apoptosis. As shown in Figure 4(a), the mitochondrial Bax level decreased in a time-dependent manner but simultaneously increased in the cytosolic fraction. The opposite was observed for the cytochrome c level. Interestingly, SP600125 markedly blocked cytochrome c release from mitochondria in SKOV3 cells exposed to allicin (Figure 4(b)). Allicin-induced JNK clearly leads directly to an increase in cytochrome c content. These biochemical changes confirm that allicin-induced apoptosis is mediated by JNK activation.


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

Xu L, Yu J, Zhai D, Zhang D, Shen W, Bai L, Cai Z, Yu C - Evid Based Complement Alternat Med (2014)

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.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4109593&req=5

fig4: 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.
Mentions: The Bax (2D2) and cytochrome c levels in the mitochondrial and cytosolic fractions were examined to further elucidate whether the JNK pathway is involved in downstream molecular events of apoptosis. As shown in Figure 4(a), the mitochondrial Bax level decreased in a time-dependent manner but simultaneously increased in the cytosolic fraction. The opposite was observed for the cytochrome c level. Interestingly, SP600125 markedly blocked cytochrome c release from mitochondria in SKOV3 cells exposed to allicin (Figure 4(b)). Allicin-induced JNK clearly leads directly to an increase in cytochrome c content. These biochemical changes confirm that allicin-induced apoptosis is mediated by JNK activation.

Bottom Line: Results.Conclusions.Our data input new insights to the literature of allicin-induced apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Traditional Chinese Gynecology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus