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Antiangiogenic and antiproliferative effects of black pomegranate peel extract on melanoma cell line.

Dana N, Javanmard ShH, Rafiee L - Res Pharm Sci (2015 Mar-Apr)

Bottom Line: VEGF mRNA expression and concentration levels in culture medium of PPE treated HUVECs reduced significantly in a concentration-dependent manner (P<0.05).Based on the results, PPE could effectively suppress angiogenesis potentially through a VEGF dependent mechanism.Further studies are needed to confirm these results.

View Article: PubMed Central - PubMed

Affiliation: Applied Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.

ABSTRACT
In the present study possible effects of black pomegranate peel extract (PPE) on the B16F10 melanoma cells proliferation and Human Umbilical Vein Endothelial Cells (HUVECs) angiogenesis were investigated. PPE was added into the cell lines (B16F10 and HUVECs) media with different concentrations (10-450 μg/ml). After 48 h, the cell survival was measured by 3-(Dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Angiogenesis was investigated by matrigel assay (PPE (200, 300, 400 μg/ml)); HUVECs, vascular endothelial growth factor (VEGF) mRNA expression was detected by quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) assay. VEGF concentration in culture medium of HUVECs was determined by enzyme-linked immunosorbent assay (ELISA). PPE had positive anti proliferative effect on melanoma cells in a dose-dependent manner, but not on HUVECs. The matrigel assay results indicated that PPE significantly inhibited length, size and junction of the tube like structures (P<0.05). VEGF mRNA expression and concentration levels in culture medium of PPE treated HUVECs reduced significantly in a concentration-dependent manner (P<0.05). Simultaneous inhibition of melanoma cell proliferation and angiogenesis proposed that, PPE can be a good candidate against melanoma development. Based on the results, PPE could effectively suppress angiogenesis potentially through a VEGF dependent mechanism. Further studies are needed to confirm these results.

No MeSH data available.


Related in: MedlinePlus

Inhibitory effect of pomegranate peel extract on tube formation of HUVECs. HUVECs were plated on the wells coated with 100 μl of Matrigel basement membrane matrix. After 24 h treatment with different doses of PPE, the cells were stained with calcein and photographed with a Nikon camera attached to a fluorescent microscope at ×10 magnification.
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Figure 2: Inhibitory effect of pomegranate peel extract on tube formation of HUVECs. HUVECs were plated on the wells coated with 100 μl of Matrigel basement membrane matrix. After 24 h treatment with different doses of PPE, the cells were stained with calcein and photographed with a Nikon camera attached to a fluorescent microscope at ×10 magnification.

Mentions: As described in earlier sections, concentrations 200, 300, and 400 μg/ml were selected to investigate the effects of PPE on angiogenesis. To ascertain the anti-angiogenic activity of PPE, the treatment started at the time of HUVEC seeding on to Matrigel. We performed the in vitro angiogenesis assay and examined the ability of endothelial cells to form tubes in the absence or presence of PPE. HUVECs tube formation was observed over a period of time. Results in Fig. 2 indicate that PPE can suppress the formation of tube-like structures at all concentrations tested in this study. Our results showed that PPE significantly reduced not only the size of the tubes (P<0.05) (Fig. 3. A) at all tested concentrations but also the length and the number of junctions of the tubes relative to the negative control group (P<0.05) (Fig.3B, 3C).


Antiangiogenic and antiproliferative effects of black pomegranate peel extract on melanoma cell line.

Dana N, Javanmard ShH, Rafiee L - Res Pharm Sci (2015 Mar-Apr)

Inhibitory effect of pomegranate peel extract on tube formation of HUVECs. HUVECs were plated on the wells coated with 100 μl of Matrigel basement membrane matrix. After 24 h treatment with different doses of PPE, the cells were stained with calcein and photographed with a Nikon camera attached to a fluorescent microscope at ×10 magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4584450&req=5

Figure 2: Inhibitory effect of pomegranate peel extract on tube formation of HUVECs. HUVECs were plated on the wells coated with 100 μl of Matrigel basement membrane matrix. After 24 h treatment with different doses of PPE, the cells were stained with calcein and photographed with a Nikon camera attached to a fluorescent microscope at ×10 magnification.
Mentions: As described in earlier sections, concentrations 200, 300, and 400 μg/ml were selected to investigate the effects of PPE on angiogenesis. To ascertain the anti-angiogenic activity of PPE, the treatment started at the time of HUVEC seeding on to Matrigel. We performed the in vitro angiogenesis assay and examined the ability of endothelial cells to form tubes in the absence or presence of PPE. HUVECs tube formation was observed over a period of time. Results in Fig. 2 indicate that PPE can suppress the formation of tube-like structures at all concentrations tested in this study. Our results showed that PPE significantly reduced not only the size of the tubes (P<0.05) (Fig. 3. A) at all tested concentrations but also the length and the number of junctions of the tubes relative to the negative control group (P<0.05) (Fig.3B, 3C).

Bottom Line: VEGF mRNA expression and concentration levels in culture medium of PPE treated HUVECs reduced significantly in a concentration-dependent manner (P<0.05).Based on the results, PPE could effectively suppress angiogenesis potentially through a VEGF dependent mechanism.Further studies are needed to confirm these results.

View Article: PubMed Central - PubMed

Affiliation: Applied Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.

ABSTRACT
In the present study possible effects of black pomegranate peel extract (PPE) on the B16F10 melanoma cells proliferation and Human Umbilical Vein Endothelial Cells (HUVECs) angiogenesis were investigated. PPE was added into the cell lines (B16F10 and HUVECs) media with different concentrations (10-450 μg/ml). After 48 h, the cell survival was measured by 3-(Dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Angiogenesis was investigated by matrigel assay (PPE (200, 300, 400 μg/ml)); HUVECs, vascular endothelial growth factor (VEGF) mRNA expression was detected by quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) assay. VEGF concentration in culture medium of HUVECs was determined by enzyme-linked immunosorbent assay (ELISA). PPE had positive anti proliferative effect on melanoma cells in a dose-dependent manner, but not on HUVECs. The matrigel assay results indicated that PPE significantly inhibited length, size and junction of the tube like structures (P<0.05). VEGF mRNA expression and concentration levels in culture medium of PPE treated HUVECs reduced significantly in a concentration-dependent manner (P<0.05). Simultaneous inhibition of melanoma cell proliferation and angiogenesis proposed that, PPE can be a good candidate against melanoma development. Based on the results, PPE could effectively suppress angiogenesis potentially through a VEGF dependent mechanism. Further studies are needed to confirm these results.

No MeSH data available.


Related in: MedlinePlus