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Role of annexin a5 on mitochondria-dependent apoptosis induced by tetramethoxystilbene in human breast cancer cells.

Hong M, Park N, Chun YJ - Biomol Ther (Seoul) (2014)

Bottom Line: Moreover, we found that the cytosolic release of apoptogenic factors such as cytochrome c, or apoptosis-inducing factor (AIF) in mitochondria was markedly increased.Annexin A5 depletion by siRNA led to decreased proapoptotic factors such as Bax, Bak, and annexin A5.Taken together, our results indicate that annexin A5 may play an important role in TMS-mediated mitochondrial apoptosis through the regulation of proapoptotic proteins and VDAC1 expression.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea.

ABSTRACT
We have previously shown that 2,4,3',5'-tetramethoxystilbene (TMS), a trans-stilbene analogue, induces apoptosis in human cancer cells. However, the detailed mechanisms of mitochondria-dependent apoptosis induced by TMS are not fully understood. In the present study, the possible roles of annexin A5 in TMS-mediated apoptosis were investigated in MCF7 human breast cancer cells. Quantitative real-time PCR analysis and Western blot analysis showed that the expression of annexin A5 was strongly increased in TMS-treated cells. TMS caused a strong translocation of annexin A5 from cytosol into mitochondria. Confocal laser scanning microscopic analysis clearly showed that TMS induced translocation of annexin A5 into mitochondria. TMS increased the expression and oligomerization of voltage-dependent anion channel (VDAC) 1, which may promote mitochondria-dependent apoptosis through disruption of mitochondrial membrane potential. When cells were treated with TMS, the levels of Bax, and Bak as well as annexin A5 were strongly enhanced. Moreover, we found that the cytosolic release of apoptogenic factors such as cytochrome c, or apoptosis-inducing factor (AIF) in mitochondria was markedly increased. Annexin A5 depletion by siRNA led to decreased proapoptotic factors such as Bax, Bak, and annexin A5. Taken together, our results indicate that annexin A5 may play an important role in TMS-mediated mitochondrial apoptosis through the regulation of proapoptotic proteins and VDAC1 expression.

No MeSH data available.


Related in: MedlinePlus

TMS regulates mitochondrial apoptotic proteins. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. After incubation, cells were harvested and the cytosolic and mitochondrial fractions were isolated. Extracted proteins were resolved by SDS-PAGE (10%) and Western blot analysis was conducted. GAPDH level was determined as loading controls for cytosolic fraction and COX-4 level was determined as a loading control for mitochondrial fraction. The data shown are representative of three independent experiments.
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f4-bt-22-519: TMS regulates mitochondrial apoptotic proteins. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. After incubation, cells were harvested and the cytosolic and mitochondrial fractions were isolated. Extracted proteins were resolved by SDS-PAGE (10%) and Western blot analysis was conducted. GAPDH level was determined as loading controls for cytosolic fraction and COX-4 level was determined as a loading control for mitochondrial fraction. The data shown are representative of three independent experiments.

Mentions: To elucidate how TMS causes mitochondria-dependent apoptosis, cells were treated with TMS (1, 5, or 10 μM) for 48 h and then the levels of Bax, Bak, cytochrome c, and AIF were measured. TMS increased mitochondrial Bax and Bak levels as well as translocation of annexin A5 into mitochondria. In addition, cytochrome c and AIF release from mitochondria to cytosol was significantly enhanced by TMS in a concentration-dependent manner (Fig. 4). These data indicated that Bax and Bak may involve in TMS-induced apoptosis through induction of cytochrome c and AIF release.


Role of annexin a5 on mitochondria-dependent apoptosis induced by tetramethoxystilbene in human breast cancer cells.

Hong M, Park N, Chun YJ - Biomol Ther (Seoul) (2014)

TMS regulates mitochondrial apoptotic proteins. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. After incubation, cells were harvested and the cytosolic and mitochondrial fractions were isolated. Extracted proteins were resolved by SDS-PAGE (10%) and Western blot analysis was conducted. GAPDH level was determined as loading controls for cytosolic fraction and COX-4 level was determined as a loading control for mitochondrial fraction. The data shown are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256031&req=5

f4-bt-22-519: TMS regulates mitochondrial apoptotic proteins. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. After incubation, cells were harvested and the cytosolic and mitochondrial fractions were isolated. Extracted proteins were resolved by SDS-PAGE (10%) and Western blot analysis was conducted. GAPDH level was determined as loading controls for cytosolic fraction and COX-4 level was determined as a loading control for mitochondrial fraction. The data shown are representative of three independent experiments.
Mentions: To elucidate how TMS causes mitochondria-dependent apoptosis, cells were treated with TMS (1, 5, or 10 μM) for 48 h and then the levels of Bax, Bak, cytochrome c, and AIF were measured. TMS increased mitochondrial Bax and Bak levels as well as translocation of annexin A5 into mitochondria. In addition, cytochrome c and AIF release from mitochondria to cytosol was significantly enhanced by TMS in a concentration-dependent manner (Fig. 4). These data indicated that Bax and Bak may involve in TMS-induced apoptosis through induction of cytochrome c and AIF release.

Bottom Line: Moreover, we found that the cytosolic release of apoptogenic factors such as cytochrome c, or apoptosis-inducing factor (AIF) in mitochondria was markedly increased.Annexin A5 depletion by siRNA led to decreased proapoptotic factors such as Bax, Bak, and annexin A5.Taken together, our results indicate that annexin A5 may play an important role in TMS-mediated mitochondrial apoptosis through the regulation of proapoptotic proteins and VDAC1 expression.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea.

ABSTRACT
We have previously shown that 2,4,3',5'-tetramethoxystilbene (TMS), a trans-stilbene analogue, induces apoptosis in human cancer cells. However, the detailed mechanisms of mitochondria-dependent apoptosis induced by TMS are not fully understood. In the present study, the possible roles of annexin A5 in TMS-mediated apoptosis were investigated in MCF7 human breast cancer cells. Quantitative real-time PCR analysis and Western blot analysis showed that the expression of annexin A5 was strongly increased in TMS-treated cells. TMS caused a strong translocation of annexin A5 from cytosol into mitochondria. Confocal laser scanning microscopic analysis clearly showed that TMS induced translocation of annexin A5 into mitochondria. TMS increased the expression and oligomerization of voltage-dependent anion channel (VDAC) 1, which may promote mitochondria-dependent apoptosis through disruption of mitochondrial membrane potential. When cells were treated with TMS, the levels of Bax, and Bak as well as annexin A5 were strongly enhanced. Moreover, we found that the cytosolic release of apoptogenic factors such as cytochrome c, or apoptosis-inducing factor (AIF) in mitochondria was markedly increased. Annexin A5 depletion by siRNA led to decreased proapoptotic factors such as Bax, Bak, and annexin A5. Taken together, our results indicate that annexin A5 may play an important role in TMS-mediated mitochondrial apoptosis through the regulation of proapoptotic proteins and VDAC1 expression.

No MeSH data available.


Related in: MedlinePlus