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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 induces annexin A5 mRNA and protein expression in MCF-7 cells. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. The mRNA and protein levels of annexin A5 were determined using quantitative real-time PCR and Western blot analysis. Total RNA was isolated and human annexin A5 genes were amplified with specific primers. Expression of GAPDH mRNA was determined as a RNA control. Total cellular lysates were prepared for Western blot analysis using annexin A5 antibody. GAPDH was used as a loading control. (A) Quantitative real-time PCR, (B) Western blot analysis. The data shown are representative of three independent experiments.
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f1-bt-22-519: TMS induces annexin A5 mRNA and protein expression in MCF-7 cells. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. The mRNA and protein levels of annexin A5 were determined using quantitative real-time PCR and Western blot analysis. Total RNA was isolated and human annexin A5 genes were amplified with specific primers. Expression of GAPDH mRNA was determined as a RNA control. Total cellular lysates were prepared for Western blot analysis using annexin A5 antibody. GAPDH was used as a loading control. (A) Quantitative real-time PCR, (B) Western blot analysis. The data shown are representative of three independent experiments.

Mentions: To elucidate whether TMS is able to induce expression of annexin A5 in MCF-7 cells, the mRNA and protein expression of annexin A5 were measured by quantitative real-time PCR and Western blot, respectively. As shown in Fig. 1A, when cells were treated with TMS (1, 5, or 10 μM) for 48 h, annexin A5 mRNA expression was significantly increased in a concentration-dependent manner. In agreement with increasing mRNA levels, annexin A5 protein level were also strongly enhanced by TMS (Fig. 1B).


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 induces annexin A5 mRNA and protein expression in MCF-7 cells. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. The mRNA and protein levels of annexin A5 were determined using quantitative real-time PCR and Western blot analysis. Total RNA was isolated and human annexin A5 genes were amplified with specific primers. Expression of GAPDH mRNA was determined as a RNA control. Total cellular lysates were prepared for Western blot analysis using annexin A5 antibody. GAPDH was used as a loading control. (A) Quantitative real-time PCR, (B) Western blot analysis. 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

f1-bt-22-519: TMS induces annexin A5 mRNA and protein expression in MCF-7 cells. MCF-7 cells were treated with various concentrations of TMS (0, 1, 5, or 10 μM) for 48 h. The mRNA and protein levels of annexin A5 were determined using quantitative real-time PCR and Western blot analysis. Total RNA was isolated and human annexin A5 genes were amplified with specific primers. Expression of GAPDH mRNA was determined as a RNA control. Total cellular lysates were prepared for Western blot analysis using annexin A5 antibody. GAPDH was used as a loading control. (A) Quantitative real-time PCR, (B) Western blot analysis. The data shown are representative of three independent experiments.
Mentions: To elucidate whether TMS is able to induce expression of annexin A5 in MCF-7 cells, the mRNA and protein expression of annexin A5 were measured by quantitative real-time PCR and Western blot, respectively. As shown in Fig. 1A, when cells were treated with TMS (1, 5, or 10 μM) for 48 h, annexin A5 mRNA expression was significantly increased in a concentration-dependent manner. In agreement with increasing mRNA levels, annexin A5 protein level were also strongly enhanced by TMS (Fig. 1B).

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