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Cortex Moutan Induces Bladder Cancer Cell Death via Apoptosis and Retards Tumor Growth in Mouse Bladders.

Lin MY, Lee YR, Chiang SY, Li YZ, Chen YS, Hsu CD, Liu YW - Evid Based Complement Alternat Med (2013)

Bottom Line: Furthermore, it has been reported that Cortex Moutan has anticancer effect.In summary, these results demonstrate the antiproliferation and anti-invasion properties of Cortex Moutan in bladder cancer cells and its antibladder tumor effect in vivo.Cortex Moutan may provide an alternative therapeutic strategy for the intravesical therapy of superficial bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Graduate Institute of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung 40402, Taiwan ; Department of Chinese Medicine, Chiayi Christian Hospital, Chiayi 60002, Taiwan.

ABSTRACT
Cortex Moutan is the root bark of Paeonia suffruticosa Andr. It is the herbal medicine widely used in Traditional Chinese Medicine for the treatment of blood-heat and blood-stasis syndrome. Furthermore, it has been reported that Cortex Moutan has anticancer effect. In this study, the Cortex Moutan extract was evaluated in bladder cancer therapy in vitro and in vivo. Cortex Moutan extract reduces cell viability with IC50 between 1~2 mg/ml in bladder cancer cells, and it has lower cytotoxicity in normal urotheliums. It arrests cells in G1 and S phase and causes phosphatidylserine expression in the outside of cell membrane. It induces caspase-8 and caspase-3 activation and poly(ADP-ribose) polymerase degradation. The pan caspase inhibitor z-VAD-fmk reverses Cortex Moutan-induced cell death. Cortex Moutan also inhibits cell invasion activity in 5637 cells. In mouse orthotopic bladder cancer model, intravesical application of Cortex Moutan decreases the bladder tumor size without altering the blood biochemical parameters. In summary, these results demonstrate the antiproliferation and anti-invasion properties of Cortex Moutan in bladder cancer cells and its antibladder tumor effect in vivo. Cortex Moutan may provide an alternative therapeutic strategy for the intravesical therapy of superficial bladder cancer.

No MeSH data available.


Related in: MedlinePlus

CM extract induces cell apoptosis. (a) Cell cycle distribution after CM treatment. Cells were treated with medium or CM extract (0, 0.5, 1, 2, 3, 3.5 mg/mL) for 24 h and 48 h then collected for cell cycle analysis. Data represent the mean ± SEM of triplicate. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with control. (b) Apoptosis analysis by Annexin-V-PI staining assay. The apoptosis percentage implied the Annexin-V-positive and PI-negative staining cells. Data represent the mean ± SEM of triplicate. ***P < 0.001 compared with control. (c) Western blot analysis of apoptotic proteins including PARP (original 116 kDa and degraded 89 kDa forms), original caspase-8, activated caspcas-3, and GAPDH (internal control). (d) z-VAD-fmk reverses CM-induced cell death. z-VAD-fmk was added in medium 1 h before CM treatment, and cell number was counted after CM treatment for 24 h. Data represent the mean ± SEM of quadruplicate. *P < 0.05, and **P < 0.01 compared with each other. All the experiments were repeated for three times.
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fig2: CM extract induces cell apoptosis. (a) Cell cycle distribution after CM treatment. Cells were treated with medium or CM extract (0, 0.5, 1, 2, 3, 3.5 mg/mL) for 24 h and 48 h then collected for cell cycle analysis. Data represent the mean ± SEM of triplicate. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with control. (b) Apoptosis analysis by Annexin-V-PI staining assay. The apoptosis percentage implied the Annexin-V-positive and PI-negative staining cells. Data represent the mean ± SEM of triplicate. ***P < 0.001 compared with control. (c) Western blot analysis of apoptotic proteins including PARP (original 116 kDa and degraded 89 kDa forms), original caspase-8, activated caspcas-3, and GAPDH (internal control). (d) z-VAD-fmk reverses CM-induced cell death. z-VAD-fmk was added in medium 1 h before CM treatment, and cell number was counted after CM treatment for 24 h. Data represent the mean ± SEM of quadruplicate. *P < 0.05, and **P < 0.01 compared with each other. All the experiments were repeated for three times.

Mentions: Cell cycle analysis was performed in MB49 and 5637 cells after CM extract treatment. CM dose dependently increased sub-G1 population while decreasing G1 and S population in MB49 and 5637 cells (Figure 2(a)). CM also increased exposed phosphatidylserine by Annex V-FITC staining assay in MB49 and 5637 cells (Figure 2(b)). The CM-induced apoptosis was also confirmed by Western Blot analysis. It shows that CM dose dependently induces the activation of caspase-8 and caspase-3, and degrades PARP in MB49 and 5637 cells (Figure 2(c)). When the pan-caspase inhibitor z-VAD-fmk was added, it partially reversed CM-induced cell death (Figure 2(d)). These results suggest that CM induces cell death at least via extrinsic apoptosis pathway, which was concurrent with the activation of caspase-8 and caspase-3.


Cortex Moutan Induces Bladder Cancer Cell Death via Apoptosis and Retards Tumor Growth in Mouse Bladders.

Lin MY, Lee YR, Chiang SY, Li YZ, Chen YS, Hsu CD, Liu YW - Evid Based Complement Alternat Med (2013)

CM extract induces cell apoptosis. (a) Cell cycle distribution after CM treatment. Cells were treated with medium or CM extract (0, 0.5, 1, 2, 3, 3.5 mg/mL) for 24 h and 48 h then collected for cell cycle analysis. Data represent the mean ± SEM of triplicate. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with control. (b) Apoptosis analysis by Annexin-V-PI staining assay. The apoptosis percentage implied the Annexin-V-positive and PI-negative staining cells. Data represent the mean ± SEM of triplicate. ***P < 0.001 compared with control. (c) Western blot analysis of apoptotic proteins including PARP (original 116 kDa and degraded 89 kDa forms), original caspase-8, activated caspcas-3, and GAPDH (internal control). (d) z-VAD-fmk reverses CM-induced cell death. z-VAD-fmk was added in medium 1 h before CM treatment, and cell number was counted after CM treatment for 24 h. Data represent the mean ± SEM of quadruplicate. *P < 0.05, and **P < 0.01 compared with each other. All the experiments were repeated for three times.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3824643&req=5

fig2: CM extract induces cell apoptosis. (a) Cell cycle distribution after CM treatment. Cells were treated with medium or CM extract (0, 0.5, 1, 2, 3, 3.5 mg/mL) for 24 h and 48 h then collected for cell cycle analysis. Data represent the mean ± SEM of triplicate. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with control. (b) Apoptosis analysis by Annexin-V-PI staining assay. The apoptosis percentage implied the Annexin-V-positive and PI-negative staining cells. Data represent the mean ± SEM of triplicate. ***P < 0.001 compared with control. (c) Western blot analysis of apoptotic proteins including PARP (original 116 kDa and degraded 89 kDa forms), original caspase-8, activated caspcas-3, and GAPDH (internal control). (d) z-VAD-fmk reverses CM-induced cell death. z-VAD-fmk was added in medium 1 h before CM treatment, and cell number was counted after CM treatment for 24 h. Data represent the mean ± SEM of quadruplicate. *P < 0.05, and **P < 0.01 compared with each other. All the experiments were repeated for three times.
Mentions: Cell cycle analysis was performed in MB49 and 5637 cells after CM extract treatment. CM dose dependently increased sub-G1 population while decreasing G1 and S population in MB49 and 5637 cells (Figure 2(a)). CM also increased exposed phosphatidylserine by Annex V-FITC staining assay in MB49 and 5637 cells (Figure 2(b)). The CM-induced apoptosis was also confirmed by Western Blot analysis. It shows that CM dose dependently induces the activation of caspase-8 and caspase-3, and degrades PARP in MB49 and 5637 cells (Figure 2(c)). When the pan-caspase inhibitor z-VAD-fmk was added, it partially reversed CM-induced cell death (Figure 2(d)). These results suggest that CM induces cell death at least via extrinsic apoptosis pathway, which was concurrent with the activation of caspase-8 and caspase-3.

Bottom Line: Furthermore, it has been reported that Cortex Moutan has anticancer effect.In summary, these results demonstrate the antiproliferation and anti-invasion properties of Cortex Moutan in bladder cancer cells and its antibladder tumor effect in vivo.Cortex Moutan may provide an alternative therapeutic strategy for the intravesical therapy of superficial bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Graduate Institute of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung 40402, Taiwan ; Department of Chinese Medicine, Chiayi Christian Hospital, Chiayi 60002, Taiwan.

ABSTRACT
Cortex Moutan is the root bark of Paeonia suffruticosa Andr. It is the herbal medicine widely used in Traditional Chinese Medicine for the treatment of blood-heat and blood-stasis syndrome. Furthermore, it has been reported that Cortex Moutan has anticancer effect. In this study, the Cortex Moutan extract was evaluated in bladder cancer therapy in vitro and in vivo. Cortex Moutan extract reduces cell viability with IC50 between 1~2 mg/ml in bladder cancer cells, and it has lower cytotoxicity in normal urotheliums. It arrests cells in G1 and S phase and causes phosphatidylserine expression in the outside of cell membrane. It induces caspase-8 and caspase-3 activation and poly(ADP-ribose) polymerase degradation. The pan caspase inhibitor z-VAD-fmk reverses Cortex Moutan-induced cell death. Cortex Moutan also inhibits cell invasion activity in 5637 cells. In mouse orthotopic bladder cancer model, intravesical application of Cortex Moutan decreases the bladder tumor size without altering the blood biochemical parameters. In summary, these results demonstrate the antiproliferation and anti-invasion properties of Cortex Moutan in bladder cancer cells and its antibladder tumor effect in vivo. Cortex Moutan may provide an alternative therapeutic strategy for the intravesical therapy of superficial bladder cancer.

No MeSH data available.


Related in: MedlinePlus