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Triggering apoptotic death of human malignant melanoma a375.s2 cells by bufalin: involvement of caspase cascade-dependent and independent mitochondrial signaling pathways.

Hsiao YP, Yu CS, Yu CC, Yang JS, Chiang JH, Lu CC, Huang HY, Tang NY, Yang JH, Huang AC, Chung JG - Evid Based Complement Alternat Med (2012)

Bottom Line: The aim of this investigation was to study the effects of bufalin on human malignant melanoma A375.S2 cells and to elucidate possible mechanisms involved in induction of apoptosis.Our results indicated that cells after exposure to bufalin significantly decreased cell viability, and induced cell morphological changes and chromatin condensation in a concentration-dependent manner.Additionally, the apoptotic induction of bufalin on A375.S2 cells resulted from mitochondrial dysfunction-related responses (disruption of the ΔΨ(m) and releases of cytochrome c, AIF, and Endo G), and activations of caspase-3, caspase-8 and caspase-9 expressions.

View Article: PubMed Central - PubMed

Affiliation: Institute of Medicine, Chung Shan Medical University, Taichung 402, Taiwan.

ABSTRACT
Bufalin was obtained from the skin and parotid venom glands of toad and has been shown to induce cytotoxic effects in various types of cancer cell lines, but there is no report to show that whether bufalin affects human skin cancer cells. The aim of this investigation was to study the effects of bufalin on human malignant melanoma A375.S2 cells and to elucidate possible mechanisms involved in induction of apoptosis. A375.S2 cells were treated with different concentrations of bufalin for a specific time period and investigated for effects on apoptotic analyses. Our results indicated that cells after exposure to bufalin significantly decreased cell viability, and induced cell morphological changes and chromatin condensation in a concentration-dependent manner. Flow cytometric assays indicated that bufalin promoted ROS productions, loss of mitochondrial membrane potential (ΔΨ(m)), intracellular Ca(2+) release, and nitric oxide (NO) formations in A375.S2 cells. Additionally, the apoptotic induction of bufalin on A375.S2 cells resulted from mitochondrial dysfunction-related responses (disruption of the ΔΨ(m) and releases of cytochrome c, AIF, and Endo G), and activations of caspase-3, caspase-8 and caspase-9 expressions. Based on those observations, we suggest that bufalin-triggered apoptosis in A375.S2 cells is correlated with extrinsic- and mitochondria-mediated multiple signal pathways.

No MeSH data available.


Related in: MedlinePlus

Bufalin induces chromatin condensation and apoptosis in A375.S2 cells. Cells were incubated with various concentrations of bufalin for 24 (a) and 48 h (b). Apoptotic cells were determined by DAPI staining and were photographed at a 200x magnification under fluorescence microscopy as described in Section 2. Each experiment was done with triple sets with similar results.
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fig2: Bufalin induces chromatin condensation and apoptosis in A375.S2 cells. Cells were incubated with various concentrations of bufalin for 24 (a) and 48 h (b). Apoptotic cells were determined by DAPI staining and were photographed at a 200x magnification under fluorescence microscopy as described in Section 2. Each experiment was done with triple sets with similar results.

Mentions: In order to confirm if bufalin-induced cell death in A375.S2 cells was through the induction of apoptosis, we used DAPI staining assay. Results shown in Figures 2(a) and 2(b) revealed that chromatin condensation and apoptotic cells were present in bufalin-treated A375.S2 cells for 24 and 48-h treatments. Moreover, the percentage of apoptotic cells is calculated (Figure 2) compared with intact control cells and this effect was concentration dependent.


Triggering apoptotic death of human malignant melanoma a375.s2 cells by bufalin: involvement of caspase cascade-dependent and independent mitochondrial signaling pathways.

Hsiao YP, Yu CS, Yu CC, Yang JS, Chiang JH, Lu CC, Huang HY, Tang NY, Yang JH, Huang AC, Chung JG - Evid Based Complement Alternat Med (2012)

Bufalin induces chromatin condensation and apoptosis in A375.S2 cells. Cells were incubated with various concentrations of bufalin for 24 (a) and 48 h (b). Apoptotic cells were determined by DAPI staining and were photographed at a 200x magnification under fluorescence microscopy as described in Section 2. Each experiment was done with triple sets with similar results.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3376545&req=5

fig2: Bufalin induces chromatin condensation and apoptosis in A375.S2 cells. Cells were incubated with various concentrations of bufalin for 24 (a) and 48 h (b). Apoptotic cells were determined by DAPI staining and were photographed at a 200x magnification under fluorescence microscopy as described in Section 2. Each experiment was done with triple sets with similar results.
Mentions: In order to confirm if bufalin-induced cell death in A375.S2 cells was through the induction of apoptosis, we used DAPI staining assay. Results shown in Figures 2(a) and 2(b) revealed that chromatin condensation and apoptotic cells were present in bufalin-treated A375.S2 cells for 24 and 48-h treatments. Moreover, the percentage of apoptotic cells is calculated (Figure 2) compared with intact control cells and this effect was concentration dependent.

Bottom Line: The aim of this investigation was to study the effects of bufalin on human malignant melanoma A375.S2 cells and to elucidate possible mechanisms involved in induction of apoptosis.Our results indicated that cells after exposure to bufalin significantly decreased cell viability, and induced cell morphological changes and chromatin condensation in a concentration-dependent manner.Additionally, the apoptotic induction of bufalin on A375.S2 cells resulted from mitochondrial dysfunction-related responses (disruption of the ΔΨ(m) and releases of cytochrome c, AIF, and Endo G), and activations of caspase-3, caspase-8 and caspase-9 expressions.

View Article: PubMed Central - PubMed

Affiliation: Institute of Medicine, Chung Shan Medical University, Taichung 402, Taiwan.

ABSTRACT
Bufalin was obtained from the skin and parotid venom glands of toad and has been shown to induce cytotoxic effects in various types of cancer cell lines, but there is no report to show that whether bufalin affects human skin cancer cells. The aim of this investigation was to study the effects of bufalin on human malignant melanoma A375.S2 cells and to elucidate possible mechanisms involved in induction of apoptosis. A375.S2 cells were treated with different concentrations of bufalin for a specific time period and investigated for effects on apoptotic analyses. Our results indicated that cells after exposure to bufalin significantly decreased cell viability, and induced cell morphological changes and chromatin condensation in a concentration-dependent manner. Flow cytometric assays indicated that bufalin promoted ROS productions, loss of mitochondrial membrane potential (ΔΨ(m)), intracellular Ca(2+) release, and nitric oxide (NO) formations in A375.S2 cells. Additionally, the apoptotic induction of bufalin on A375.S2 cells resulted from mitochondrial dysfunction-related responses (disruption of the ΔΨ(m) and releases of cytochrome c, AIF, and Endo G), and activations of caspase-3, caspase-8 and caspase-9 expressions. Based on those observations, we suggest that bufalin-triggered apoptosis in A375.S2 cells is correlated with extrinsic- and mitochondria-mediated multiple signal pathways.

No MeSH data available.


Related in: MedlinePlus