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Esculetin, a Coumarin Derivative, Exhibits Anti-proliferative and Pro-apoptotic Activity in G361 Human Malignant Melanoma.

Jeon YJ, Jang JY, Shim JH, Myung PK, Chae JI - J Cancer Prev (2015)

Bottom Line: Esculetin exhibited significant anti-proliferative effects on the HMM cells in a dose-dependent manner.Interestingly, we found that esculetin induced nuclear shrinkage and fragmentation, typical apoptosis markers, by suppression of Sp1 transcription factor (Sp1).Our results clearly demonstrated that esculetin induced apoptosis in the HMM cells by downregulating Sp1 protein levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Dental Pharmacology, School of Dentistry and Institute of Oral Bioscience, BK21 Plus, Chonbuk National University, Jeonju, Chungnam National University, Daejeon, Korea.

ABSTRACT

Background: Although esculetin, a coumarin compound, is known to induce apoptosis in human cancer cells, the effects and molecular mechanisms on the apoptosis in human malignant melanoma (HMM) cells are not well understood yet. In this study, we investigated the anti-proliferative effects of esculetin on the G361 HMM cells.

Methods: We analyzed the anti-proliferative effects and molecular mechanisms of esculetin on G361 cells by a 3-(4,5-dimethylthiazol- 2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, 4',6-diamidino-2-phenylindole staining and Western blotting.

Results: Esculetin exhibited significant anti-proliferative effects on the HMM cells in a dose-dependent manner. Interestingly, we found that esculetin induced nuclear shrinkage and fragmentation, typical apoptosis markers, by suppression of Sp1 transcription factor (Sp1). Notably, esculetin modulated Sp1 downstream target genes including p27, p21 and cyclin D1, resulted in activation of apoptosis signaling molecules such as caspase-3 and PARP in G361 HMM cells.

Conclusions: Our results clearly demonstrated that esculetin induced apoptosis in the HMM cells by downregulating Sp1 protein levels. Thus, we suggest that esculetin may be a potential anti-proliferative agent that induces apoptotic cell death in G361 HMM cells.

No MeSH data available.


Related in: MedlinePlus

The effect of esculetin on the expression of cell cycle arrest- and apoptosis-related proteins in human malignant melanoma. (A) The G361 cells were incubated with esculetin (20, 40, and 80 μg/mL) or without for 48 hours. The cell lysates were determined using Western blot analysis with antibodies against p27, p21, and cyclin D1. Equal loading protein was confirmed using glyceraldehyde-3-phosphate de-hydrogenase (GAPDH). (B) The G361 cells were treated with esculetin (0, 20, 40, and 80 μg/mL) for 48 hours. The cell lysates were then determined via Western blot analysis with antibodies against procaspase 3, active caspase 3, Bax, and PARP. Equal loading protein was confirmed using GAPDH. The values were measured using Image J densitometry, and the data were expressed as a representative of two or three independent experiments.
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f4-jcp-20-106: The effect of esculetin on the expression of cell cycle arrest- and apoptosis-related proteins in human malignant melanoma. (A) The G361 cells were incubated with esculetin (20, 40, and 80 μg/mL) or without for 48 hours. The cell lysates were determined using Western blot analysis with antibodies against p27, p21, and cyclin D1. Equal loading protein was confirmed using glyceraldehyde-3-phosphate de-hydrogenase (GAPDH). (B) The G361 cells were treated with esculetin (0, 20, 40, and 80 μg/mL) for 48 hours. The cell lysates were then determined via Western blot analysis with antibodies against procaspase 3, active caspase 3, Bax, and PARP. Equal loading protein was confirmed using GAPDH. The values were measured using Image J densitometry, and the data were expressed as a representative of two or three independent experiments.

Mentions: The treatment of cells with esculetin regulated the expression levels of several cell cycle related proteins (p27, p21, and cyclin D1) (Fig. 4A) and apoptosis related proteins (procaspase 3, active caspase 3, Bax, and PARP) (Fig. 4B). The G361 cells were treated with different doses of esculetin for 48 hours and were harvested. The protein expression levels of p27, p21, cyclin D1, procaspase 3, active caspase 3, Bax, and PARP were analyzed by Western blotting. The results indicate that esculetin increased p21, p27, Bax and active caspase 3 protein levels and decreased cyclinD1, procaspase 3, and PARP protein levels in the G361 cells. These results suggest that esculetin treatment in G361 cells suppresses Sp1 protein levels, resulting in cell cycle arrest and apoptotic cell death.


Esculetin, a Coumarin Derivative, Exhibits Anti-proliferative and Pro-apoptotic Activity in G361 Human Malignant Melanoma.

Jeon YJ, Jang JY, Shim JH, Myung PK, Chae JI - J Cancer Prev (2015)

The effect of esculetin on the expression of cell cycle arrest- and apoptosis-related proteins in human malignant melanoma. (A) The G361 cells were incubated with esculetin (20, 40, and 80 μg/mL) or without for 48 hours. The cell lysates were determined using Western blot analysis with antibodies against p27, p21, and cyclin D1. Equal loading protein was confirmed using glyceraldehyde-3-phosphate de-hydrogenase (GAPDH). (B) The G361 cells were treated with esculetin (0, 20, 40, and 80 μg/mL) for 48 hours. The cell lysates were then determined via Western blot analysis with antibodies against procaspase 3, active caspase 3, Bax, and PARP. Equal loading protein was confirmed using GAPDH. The values were measured using Image J densitometry, and the data were expressed as a representative of two or three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492354&req=5

f4-jcp-20-106: The effect of esculetin on the expression of cell cycle arrest- and apoptosis-related proteins in human malignant melanoma. (A) The G361 cells were incubated with esculetin (20, 40, and 80 μg/mL) or without for 48 hours. The cell lysates were determined using Western blot analysis with antibodies against p27, p21, and cyclin D1. Equal loading protein was confirmed using glyceraldehyde-3-phosphate de-hydrogenase (GAPDH). (B) The G361 cells were treated with esculetin (0, 20, 40, and 80 μg/mL) for 48 hours. The cell lysates were then determined via Western blot analysis with antibodies against procaspase 3, active caspase 3, Bax, and PARP. Equal loading protein was confirmed using GAPDH. The values were measured using Image J densitometry, and the data were expressed as a representative of two or three independent experiments.
Mentions: The treatment of cells with esculetin regulated the expression levels of several cell cycle related proteins (p27, p21, and cyclin D1) (Fig. 4A) and apoptosis related proteins (procaspase 3, active caspase 3, Bax, and PARP) (Fig. 4B). The G361 cells were treated with different doses of esculetin for 48 hours and were harvested. The protein expression levels of p27, p21, cyclin D1, procaspase 3, active caspase 3, Bax, and PARP were analyzed by Western blotting. The results indicate that esculetin increased p21, p27, Bax and active caspase 3 protein levels and decreased cyclinD1, procaspase 3, and PARP protein levels in the G361 cells. These results suggest that esculetin treatment in G361 cells suppresses Sp1 protein levels, resulting in cell cycle arrest and apoptotic cell death.

Bottom Line: Esculetin exhibited significant anti-proliferative effects on the HMM cells in a dose-dependent manner.Interestingly, we found that esculetin induced nuclear shrinkage and fragmentation, typical apoptosis markers, by suppression of Sp1 transcription factor (Sp1).Our results clearly demonstrated that esculetin induced apoptosis in the HMM cells by downregulating Sp1 protein levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Dental Pharmacology, School of Dentistry and Institute of Oral Bioscience, BK21 Plus, Chonbuk National University, Jeonju, Chungnam National University, Daejeon, Korea.

ABSTRACT

Background: Although esculetin, a coumarin compound, is known to induce apoptosis in human cancer cells, the effects and molecular mechanisms on the apoptosis in human malignant melanoma (HMM) cells are not well understood yet. In this study, we investigated the anti-proliferative effects of esculetin on the G361 HMM cells.

Methods: We analyzed the anti-proliferative effects and molecular mechanisms of esculetin on G361 cells by a 3-(4,5-dimethylthiazol- 2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, 4',6-diamidino-2-phenylindole staining and Western blotting.

Results: Esculetin exhibited significant anti-proliferative effects on the HMM cells in a dose-dependent manner. Interestingly, we found that esculetin induced nuclear shrinkage and fragmentation, typical apoptosis markers, by suppression of Sp1 transcription factor (Sp1). Notably, esculetin modulated Sp1 downstream target genes including p27, p21 and cyclin D1, resulted in activation of apoptosis signaling molecules such as caspase-3 and PARP in G361 HMM cells.

Conclusions: Our results clearly demonstrated that esculetin induced apoptosis in the HMM cells by downregulating Sp1 protein levels. Thus, we suggest that esculetin may be a potential anti-proliferative agent that induces apoptotic cell death in G361 HMM cells.

No MeSH data available.


Related in: MedlinePlus