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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 disrupts bFGF-induced angiogenesis in vivo.PO and bFGF were loaded on CAMs of 10-day-old fertilized chicken eggs. After 72 h incubation, a fat emulsion was injected into the CAMs for better visualization of the blood vessels. Thermanox disc and surrounding CAMs were photographed. (A) Untreated control, (B) bFGF-treated control, (C) 0.25 µg of PO with bFGF, and (D) 0.5 µg of PO with bFGF. (E) The number of newly formed blood vessels per field was counted. All data are presented as mean ± S.D., n = 5. Statistically significant differences between control and sample groups were calculated by Student's t-test. ##, p<0.05 versus untreated control.*, p<0.05 and ***, p<0.001 versus bFGF control.
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pone-0012358-g004: PO disrupts bFGF-induced angiogenesis in vivo.PO and bFGF were loaded on CAMs of 10-day-old fertilized chicken eggs. After 72 h incubation, a fat emulsion was injected into the CAMs for better visualization of the blood vessels. Thermanox disc and surrounding CAMs were photographed. (A) Untreated control, (B) bFGF-treated control, (C) 0.25 µg of PO with bFGF, and (D) 0.5 µg of PO with bFGF. (E) The number of newly formed blood vessels per field was counted. All data are presented as mean ± S.D., n = 5. Statistically significant differences between control and sample groups were calculated by Student's t-test. ##, p<0.05 versus untreated control.*, p<0.05 and ***, p<0.001 versus bFGF control.

Mentions: To confirm the anti-angiogenic potential of PO, chick CAM assay was employed in the physiological context of vascular endothelial cells within intact vessels. As shown in Fig. 4, the treatment with bFGF (100 ng/embryo) resulted in a 2.6-fold increase of new blood vessels under the thermonox disc applied on the CAMs. Furthermore, PO treatment at both doses of 0.25 and 0.5 µg/egg almost totally ified bFGF-induced angiogenesis without apparent thrombosis and hemorrhage.


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 disrupts bFGF-induced angiogenesis in vivo.PO and bFGF were loaded on CAMs of 10-day-old fertilized chicken eggs. After 72 h incubation, a fat emulsion was injected into the CAMs for better visualization of the blood vessels. Thermanox disc and surrounding CAMs were photographed. (A) Untreated control, (B) bFGF-treated control, (C) 0.25 µg of PO with bFGF, and (D) 0.5 µg of PO with bFGF. (E) The number of newly formed blood vessels per field was counted. All data are presented as mean ± S.D., n = 5. Statistically significant differences between control and sample groups were calculated by Student's t-test. ##, p<0.05 versus untreated control.*, p<0.05 and ***, p<0.001 versus bFGF control.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0012358-g004: PO disrupts bFGF-induced angiogenesis in vivo.PO and bFGF were loaded on CAMs of 10-day-old fertilized chicken eggs. After 72 h incubation, a fat emulsion was injected into the CAMs for better visualization of the blood vessels. Thermanox disc and surrounding CAMs were photographed. (A) Untreated control, (B) bFGF-treated control, (C) 0.25 µg of PO with bFGF, and (D) 0.5 µg of PO with bFGF. (E) The number of newly formed blood vessels per field was counted. All data are presented as mean ± S.D., n = 5. Statistically significant differences between control and sample groups were calculated by Student's t-test. ##, p<0.05 versus untreated control.*, p<0.05 and ***, p<0.001 versus bFGF control.
Mentions: To confirm the anti-angiogenic potential of PO, chick CAM assay was employed in the physiological context of vascular endothelial cells within intact vessels. As shown in Fig. 4, the treatment with bFGF (100 ng/embryo) resulted in a 2.6-fold increase of new blood vessels under the thermonox disc applied on the CAMs. Furthermore, PO treatment at both doses of 0.25 and 0.5 µg/egg almost totally ified bFGF-induced angiogenesis without apparent thrombosis and hemorrhage.

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