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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

Apoptotic effect induced by esculetin in human malignant melanoma cells. The G361 cells were incubated with esculetin (0 and 80 μg/mL) for 48 hours. (A) Fluorescence microscopy images of the DAPI-stained cells. The white arrows represent DNA fragmentation and chromatin condensation. Scale bars: 20 μm. (B) The DNA fragmentation and chromatin condensation were determined to have a significant difference relative to untreated control cells, as indicated by an asterisk (*P < 0.05).
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f2-jcp-20-106: Apoptotic effect induced by esculetin in human malignant melanoma cells. The G361 cells were incubated with esculetin (0 and 80 μg/mL) for 48 hours. (A) Fluorescence microscopy images of the DAPI-stained cells. The white arrows represent DNA fragmentation and chromatin condensation. Scale bars: 20 μm. (B) The DNA fragmentation and chromatin condensation were determined to have a significant difference relative to untreated control cells, as indicated by an asterisk (*P < 0.05).

Mentions: The morphological changes including cytoplasmic blebbing, irregular cell morphology change and detachment from cell culture vessels are critical hallmarkers of apoptosis.26 To determine whether esculetin induced apoptosis or not, we examined the nuclei integrity by esculetin treatment. G361 HMM cells were treated with 60 μg/mL of esculetin for 48 hours and stained with DAPI. We found that esculatin dramatically induced nuclei condensation and fragmentation compared with dimethyl sulfoxide treated control cells (Fig. 2). These results demonstrated that esculetin suppressed cell proliferation by induction of apoptosis.


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)

Apoptotic effect induced by esculetin in human malignant melanoma cells. The G361 cells were incubated with esculetin (0 and 80 μg/mL) for 48 hours. (A) Fluorescence microscopy images of the DAPI-stained cells. The white arrows represent DNA fragmentation and chromatin condensation. Scale bars: 20 μm. (B) The DNA fragmentation and chromatin condensation were determined to have a significant difference relative to untreated control cells, as indicated by an asterisk (*P < 0.05).
© Copyright Policy
Related In: Results  -  Collection

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

f2-jcp-20-106: Apoptotic effect induced by esculetin in human malignant melanoma cells. The G361 cells were incubated with esculetin (0 and 80 μg/mL) for 48 hours. (A) Fluorescence microscopy images of the DAPI-stained cells. The white arrows represent DNA fragmentation and chromatin condensation. Scale bars: 20 μm. (B) The DNA fragmentation and chromatin condensation were determined to have a significant difference relative to untreated control cells, as indicated by an asterisk (*P < 0.05).
Mentions: The morphological changes including cytoplasmic blebbing, irregular cell morphology change and detachment from cell culture vessels are critical hallmarkers of apoptosis.26 To determine whether esculetin induced apoptosis or not, we examined the nuclei integrity by esculetin treatment. G361 HMM cells were treated with 60 μg/mL of esculetin for 48 hours and stained with DAPI. We found that esculatin dramatically induced nuclei condensation and fragmentation compared with dimethyl sulfoxide treated control cells (Fig. 2). These results demonstrated that esculetin suppressed cell proliferation by induction of apoptosis.

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