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Paeonol oxime inhibits bFGF-induced angiogenesis and reduces VEGF levels in fibrosarcoma cells.

Lee HJ, Kim SA, Lee HJ, Jeong SJ, Han I, Jung JH, Lee EO, Zhu S, Chen CY, Kim SH - PLoS ONE (2010)

Bottom Line: The treatment with PO at 12.5 microg/ml reduced the levels of phosphorylated AKT and VEGF expression (intracellular and extracelluar) in HT-1080 cells.Consistently, immunefluorescence imaging analysis revealed that PO treatment attenuated AKT phosphorylation in HT-1080 cells.Taken together, these results suggest that PO inhibits bFGF-induced angiogenesis in HUVECs and decreased the levels of PI3K, phospho-AKT and VEGF in HT-1080 cells.

View Article: PubMed Central - PubMed

Affiliation: College of Oriental Medicine, Kyung Hee University, Seoul, Republic of Korea.

ABSTRACT

Background: We previously reported the anti-angiogenic activity of paeonol isolated from Moutan Cortex. In the present study, we investigated the negative effect of paeonol oxime (PO, a paeonol derivative) on basic fibroblast growth factor (bFGF)-mediated angiogenesis in human umbilical vein endothelial cells (HUVECs) (including tumor angiogenesis) and pro-survival activity in HT-1080 fibrosarcoma cell line.

Methodology/principal findings: We showed that PO (IC(50) = 17.3 microg/ml) significantly inhibited bFGF-induced cell proliferation, which was achieved with higher concentrations of paeonol (IC(50) over 200 microg). The treatment with PO blocked bFGF-stimulated migration and in vitro capillary differentiation (tube formation) in a dose-dependent manner. Furthermore, PO was able to disrupt neovascularization in vivo. Interestingly, PO (25 microg/ml) decreased the cell viability of HT-1080 fibrosarcoma cells but not that of HUVECs. The treatment with PO at 12.5 microg/ml reduced the levels of phosphorylated AKT and VEGF expression (intracellular and extracelluar) in HT-1080 cells. Consistently, immunefluorescence imaging analysis revealed that PO treatment attenuated AKT phosphorylation in HT-1080 cells.

Conclusions/significance: Taken together, these results suggest that PO inhibits bFGF-induced angiogenesis in HUVECs and decreased the levels of PI3K, phospho-AKT and VEGF in HT-1080 cells.

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Related in: MedlinePlus

PO inhibits bFGF-induced migration and tube formation of HUVECs.Cell migration through gelatin-coated filters was measured by using the Boyden chamber. HUVECs (5×104 cells/well) were plated into the upper chamber with or without various concentrations of PO and incubated for 6 h at 37°C in a 5% CO2 incubator. Cells migrated to the lower surface were photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. (B) Tube formation assay was performed using growth factor reduced Matrigel. Cells were fixed with Diff-Quick solution, photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. All data were expressed as mean ± S.D. The statistically significant differences between control and PO treated groups were calculated by the Student's t-test. ###, p<0.001 versus untreated control.*, p<0.05 and **, p<0.01 versus bFGF control.
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pone-0012358-g003: PO inhibits bFGF-induced migration and tube formation of HUVECs.Cell migration through gelatin-coated filters was measured by using the Boyden chamber. HUVECs (5×104 cells/well) were plated into the upper chamber with or without various concentrations of PO and incubated for 6 h at 37°C in a 5% CO2 incubator. Cells migrated to the lower surface were photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. (B) Tube formation assay was performed using growth factor reduced Matrigel. Cells were fixed with Diff-Quick solution, photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. All data were expressed as mean ± S.D. The statistically significant differences between control and PO treated groups were calculated by the Student's t-test. ###, p<0.001 versus untreated control.*, p<0.05 and **, p<0.01 versus bFGF control.

Mentions: To explore the effects of PO on bFGF-stimulated angiogenesis, the migration assay using the Boyden chamber was performed. As shown in Fig. 3A, cell motility of HUVECs was increased by 10.9-fold after bFGF treatment as compared with untreated control. In contrast, PO significantly inhibited bFGF-induced migration of HUVECs, in a concentration-dependent manner with IC50 of 5.3 µg/ml.


Paeonol oxime inhibits bFGF-induced angiogenesis and reduces VEGF levels in fibrosarcoma cells.

Lee HJ, Kim SA, Lee HJ, Jeong SJ, Han I, Jung JH, Lee EO, Zhu S, Chen CY, Kim SH - PLoS ONE (2010)

PO inhibits bFGF-induced migration and tube formation of HUVECs.Cell migration through gelatin-coated filters was measured by using the Boyden chamber. HUVECs (5×104 cells/well) were plated into the upper chamber with or without various concentrations of PO and incubated for 6 h at 37°C in a 5% CO2 incubator. Cells migrated to the lower surface were photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. (B) Tube formation assay was performed using growth factor reduced Matrigel. Cells were fixed with Diff-Quick solution, photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. All data were expressed as mean ± S.D. The statistically significant differences between control and PO treated groups were calculated by the Student's t-test. ###, p<0.001 versus untreated control.*, p<0.05 and **, p<0.01 versus bFGF control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2925949&req=5

pone-0012358-g003: PO inhibits bFGF-induced migration and tube formation of HUVECs.Cell migration through gelatin-coated filters was measured by using the Boyden chamber. HUVECs (5×104 cells/well) were plated into the upper chamber with or without various concentrations of PO and incubated for 6 h at 37°C in a 5% CO2 incubator. Cells migrated to the lower surface were photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. (B) Tube formation assay was performed using growth factor reduced Matrigel. Cells were fixed with Diff-Quick solution, photographed randomly under an Axiovert S 100 light microscope at ×100 magnification and counted. All data were expressed as mean ± S.D. The statistically significant differences between control and PO treated groups were calculated by the Student's t-test. ###, p<0.001 versus untreated control.*, p<0.05 and **, p<0.01 versus bFGF control.
Mentions: To explore the effects of PO on bFGF-stimulated angiogenesis, the migration assay using the Boyden chamber was performed. As shown in Fig. 3A, cell motility of HUVECs was increased by 10.9-fold after bFGF treatment as compared with untreated control. In contrast, PO significantly inhibited bFGF-induced migration of HUVECs, in a concentration-dependent manner with IC50 of 5.3 µg/ml.

Bottom Line: The treatment with PO at 12.5 microg/ml reduced the levels of phosphorylated AKT and VEGF expression (intracellular and extracelluar) in HT-1080 cells.Consistently, immunefluorescence imaging analysis revealed that PO treatment attenuated AKT phosphorylation in HT-1080 cells.Taken together, these results suggest that PO inhibits bFGF-induced angiogenesis in HUVECs and decreased the levels of PI3K, phospho-AKT and VEGF in HT-1080 cells.

View Article: PubMed Central - PubMed

Affiliation: College of Oriental Medicine, Kyung Hee University, Seoul, Republic of Korea.

ABSTRACT

Background: We previously reported the anti-angiogenic activity of paeonol isolated from Moutan Cortex. In the present study, we investigated the negative effect of paeonol oxime (PO, a paeonol derivative) on basic fibroblast growth factor (bFGF)-mediated angiogenesis in human umbilical vein endothelial cells (HUVECs) (including tumor angiogenesis) and pro-survival activity in HT-1080 fibrosarcoma cell line.

Methodology/principal findings: We showed that PO (IC(50) = 17.3 microg/ml) significantly inhibited bFGF-induced cell proliferation, which was achieved with higher concentrations of paeonol (IC(50) over 200 microg). The treatment with PO blocked bFGF-stimulated migration and in vitro capillary differentiation (tube formation) in a dose-dependent manner. Furthermore, PO was able to disrupt neovascularization in vivo. Interestingly, PO (25 microg/ml) decreased the cell viability of HT-1080 fibrosarcoma cells but not that of HUVECs. The treatment with PO at 12.5 microg/ml reduced the levels of phosphorylated AKT and VEGF expression (intracellular and extracelluar) in HT-1080 cells. Consistently, immunefluorescence imaging analysis revealed that PO treatment attenuated AKT phosphorylation in HT-1080 cells.

Conclusions/significance: Taken together, these results suggest that PO inhibits bFGF-induced angiogenesis in HUVECs and decreased the levels of PI3K, phospho-AKT and VEGF in HT-1080 cells.

Show MeSH
Related in: MedlinePlus