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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

Quantitative analysis of mRNA expression of VEGF in HUVEC by Quantitative real time RT-PCR (Taqman). Total RNA were extracted from HUVEC in medium containing different treatments. The relative expression of VEGF were normalized to GAPDH levels measured in the same RNA preparation. Data shown are from three independent experiments analyzed in duplicate.
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Figure 4: Quantitative analysis of mRNA expression of VEGF in HUVEC by Quantitative real time RT-PCR (Taqman). Total RNA were extracted from HUVEC in medium containing different treatments. The relative expression of VEGF were normalized to GAPDH levels measured in the same RNA preparation. Data shown are from three independent experiments analyzed in duplicate.

Mentions: After optimization of the QRT-PCR, expression of VEGF and GAPDH genes was determined using Quantitative real-time RT-PCR in the treated and control cells. Relative expression of VEGF gene was determined by dividing its expression amount to that of the GAPDH gene. There was a concentration-dependent relationship between PPE and its inhibitory effect on VEGF gene expression at 200, 300, and 400 μg/ml and the concentrations of 300 and 400 μg/ml significantly inhibited VEGF mRNA expression (P<0.05) (Fig. 4).


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)

Quantitative analysis of mRNA expression of VEGF in HUVEC by Quantitative real time RT-PCR (Taqman). Total RNA were extracted from HUVEC in medium containing different treatments. The relative expression of VEGF were normalized to GAPDH levels measured in the same RNA preparation. Data shown are from three independent experiments analyzed in duplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Quantitative analysis of mRNA expression of VEGF in HUVEC by Quantitative real time RT-PCR (Taqman). Total RNA were extracted from HUVEC in medium containing different treatments. The relative expression of VEGF were normalized to GAPDH levels measured in the same RNA preparation. Data shown are from three independent experiments analyzed in duplicate.
Mentions: After optimization of the QRT-PCR, expression of VEGF and GAPDH genes was determined using Quantitative real-time RT-PCR in the treated and control cells. Relative expression of VEGF gene was determined by dividing its expression amount to that of the GAPDH gene. There was a concentration-dependent relationship between PPE and its inhibitory effect on VEGF gene expression at 200, 300, and 400 μg/ml and the concentrations of 300 and 400 μg/ml significantly inhibited VEGF mRNA expression (P<0.05) (Fig. 4).

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