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An evaluation of the anti-angiogenic effect of the Korean medicinal formula "Sa-mi-yeon-geon-tang" in vitro and in ovo.

Yi JM, Bang OS, Kim NS - BMC Complement Altern Med (2015)

Bottom Line: These anti-angiogenic effects of SMYGT in HUVECs were related to decreases in the phosphorylation of focal adhesion kinase and the expression of matrix metallopeptidase-2 activity.SMYGT exhibited an anti-angiogenic potential in both in vitro and in ovo experiments, which may partially contribute to its anti-tumor effect in clinical conditions.We suggest that SMYGT may be a promising source material for the development of anti-cancer chemotherapeutics that target angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: KM-Based Herbal Drug Development Group, Korea Institute of Oriental Medicine, 305-811, Daejeon, Republic of Korea. jmyi@kiom.re.kr.

ABSTRACT

Background: Angiogenesis is a general hallmark of cancer; therefore, the inhibition of tumor-derived angiogenesis is considered to be an attractive target in the development of anti-cancer agents. Sa-mi-yeon-geon-tang (SMYGT), a decoction that consists of four natural medicinal products, has been traditionally prescribed in Oriental medicine to treat diverse diseases, including cancer. In the present study, we investigated the anti-angiogenic potential of SMYGT in vitro and in ovo.

Methods: The anti-angiogenic potential of SMYGT was evaluated using conventional in vitro assays with human umbilical vein endothelial cells (HUVECs) and chorioallantoic membrane (CAM) assays with fertilized eggs. The expression changes of pro-angiogenic proteins and intracellular signaling in HUVECs following SMYGT treatment were determined by quantitative polymerase chain reaction, gelatinase zymography, and western blot analysis.

Results: SMYGT efficiently inhibited three-dimensional capillary-like tube formation by HUVECs on extracellular matrix supports, as well as new vessel formation on CAMs. SMYGT inhibited cell adhesion to the extracellular matrix and HUVEC cell invasion through Matrigel without affecting cell proliferation, viability, and motility. These anti-angiogenic effects of SMYGT in HUVECs were related to decreases in the phosphorylation of focal adhesion kinase and the expression of matrix metallopeptidase-2 activity.

Conclusions: SMYGT exhibited an anti-angiogenic potential in both in vitro and in ovo experiments, which may partially contribute to its anti-tumor effect in clinical conditions. We suggest that SMYGT may be a promising source material for the development of anti-cancer chemotherapeutics that target angiogenesis.

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The effects of SMYGT on FAK signaling and MMP2 expression in HUVECs. (A) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent effect of SMYGT on phosphorylation of the intracellular FAK protein were determined using western blot analyses. β-actin was used to confirm equal loading of total proteins. (B) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent changes of MMP2 enzyme activities and mRNA induced by SMYGT were determined by zymography (top) and qPCR (bottom), respectively. A PAGE gel stained with Coomassie blue solution (middle) is presented to demonstrate equal loading of the proteins for the MMP2 zymography. V, vehicle; S, SMYGT.
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Fig3: The effects of SMYGT on FAK signaling and MMP2 expression in HUVECs. (A) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent effect of SMYGT on phosphorylation of the intracellular FAK protein were determined using western blot analyses. β-actin was used to confirm equal loading of total proteins. (B) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent changes of MMP2 enzyme activities and mRNA induced by SMYGT were determined by zymography (top) and qPCR (bottom), respectively. A PAGE gel stained with Coomassie blue solution (middle) is presented to demonstrate equal loading of the proteins for the MMP2 zymography. V, vehicle; S, SMYGT.

Mentions: As shown in Figure 2C, SMYGT potently inhibited the adhesion of HUVECs to matrix supports. Because integrin-FAK signaling plays a key role in cell adhesion, and activation of integrin leads to phosphorylation of FAK [12], we determined the level of phosphorylated FAK following SMYGT treatment in HUVECs. SMYGT reduced the phosphorylation of FAK compared to vehicle treatment (Figure 3A). The total FAK and β-actin, a loading control for western blot analysis, were not changed by SMYGT treatment. Therefore, SMYGT may affect cell adhesion by down-regulating integrin-FAK signaling pathway.Figure 3


An evaluation of the anti-angiogenic effect of the Korean medicinal formula "Sa-mi-yeon-geon-tang" in vitro and in ovo.

Yi JM, Bang OS, Kim NS - BMC Complement Altern Med (2015)

The effects of SMYGT on FAK signaling and MMP2 expression in HUVECs. (A) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent effect of SMYGT on phosphorylation of the intracellular FAK protein were determined using western blot analyses. β-actin was used to confirm equal loading of total proteins. (B) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent changes of MMP2 enzyme activities and mRNA induced by SMYGT were determined by zymography (top) and qPCR (bottom), respectively. A PAGE gel stained with Coomassie blue solution (middle) is presented to demonstrate equal loading of the proteins for the MMP2 zymography. V, vehicle; S, SMYGT.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359561&req=5

Fig3: The effects of SMYGT on FAK signaling and MMP2 expression in HUVECs. (A) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent effect of SMYGT on phosphorylation of the intracellular FAK protein were determined using western blot analyses. β-actin was used to confirm equal loading of total proteins. (B) Time-(left, 200 μg/mL of SMYGT) and dose-(right, 24 h) dependent changes of MMP2 enzyme activities and mRNA induced by SMYGT were determined by zymography (top) and qPCR (bottom), respectively. A PAGE gel stained with Coomassie blue solution (middle) is presented to demonstrate equal loading of the proteins for the MMP2 zymography. V, vehicle; S, SMYGT.
Mentions: As shown in Figure 2C, SMYGT potently inhibited the adhesion of HUVECs to matrix supports. Because integrin-FAK signaling plays a key role in cell adhesion, and activation of integrin leads to phosphorylation of FAK [12], we determined the level of phosphorylated FAK following SMYGT treatment in HUVECs. SMYGT reduced the phosphorylation of FAK compared to vehicle treatment (Figure 3A). The total FAK and β-actin, a loading control for western blot analysis, were not changed by SMYGT treatment. Therefore, SMYGT may affect cell adhesion by down-regulating integrin-FAK signaling pathway.Figure 3

Bottom Line: These anti-angiogenic effects of SMYGT in HUVECs were related to decreases in the phosphorylation of focal adhesion kinase and the expression of matrix metallopeptidase-2 activity.SMYGT exhibited an anti-angiogenic potential in both in vitro and in ovo experiments, which may partially contribute to its anti-tumor effect in clinical conditions.We suggest that SMYGT may be a promising source material for the development of anti-cancer chemotherapeutics that target angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: KM-Based Herbal Drug Development Group, Korea Institute of Oriental Medicine, 305-811, Daejeon, Republic of Korea. jmyi@kiom.re.kr.

ABSTRACT

Background: Angiogenesis is a general hallmark of cancer; therefore, the inhibition of tumor-derived angiogenesis is considered to be an attractive target in the development of anti-cancer agents. Sa-mi-yeon-geon-tang (SMYGT), a decoction that consists of four natural medicinal products, has been traditionally prescribed in Oriental medicine to treat diverse diseases, including cancer. In the present study, we investigated the anti-angiogenic potential of SMYGT in vitro and in ovo.

Methods: The anti-angiogenic potential of SMYGT was evaluated using conventional in vitro assays with human umbilical vein endothelial cells (HUVECs) and chorioallantoic membrane (CAM) assays with fertilized eggs. The expression changes of pro-angiogenic proteins and intracellular signaling in HUVECs following SMYGT treatment were determined by quantitative polymerase chain reaction, gelatinase zymography, and western blot analysis.

Results: SMYGT efficiently inhibited three-dimensional capillary-like tube formation by HUVECs on extracellular matrix supports, as well as new vessel formation on CAMs. SMYGT inhibited cell adhesion to the extracellular matrix and HUVEC cell invasion through Matrigel without affecting cell proliferation, viability, and motility. These anti-angiogenic effects of SMYGT in HUVECs were related to decreases in the phosphorylation of focal adhesion kinase and the expression of matrix metallopeptidase-2 activity.

Conclusions: SMYGT exhibited an anti-angiogenic potential in both in vitro and in ovo experiments, which may partially contribute to its anti-tumor effect in clinical conditions. We suggest that SMYGT may be a promising source material for the development of anti-cancer chemotherapeutics that target angiogenesis.

Show MeSH
Related in: MedlinePlus