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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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Anti-angiogenic potential of SMYGT in vitro and in ovo. (A) HUVECs cultured on BME-pre-coated supports were exposed to different concentrations of SMYGT for 12 h. The tube length and branch numbers were determined by analyzing digitally captured images. Sulforaphane (Sulfo, 5 μM) was administered in parallel as a positive control drug. Relative tube formation was determined by comparing each group with the vehicle treated group (0 μg/mL SMYGT), and the data are presented as the means ± standard deviation (S.D.) of triplicate experiments. *P < 0.05, **P < 0.01, ***P < 0.001. (B) At ED4.5, A quarter size of thermanox coverslips containing variable amounts of SMYGT (0, 200, 400 μg/disc) were applied to the CAMs. After 2 days of incubation, 10% skimmed milk solution was injected into the CAM for observation of the inhibition zone of angiogenesis and digital images were captured. Retinoic acid (RA, 1 μg/disc) used as a control for inhibition of new vessel formation.
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Fig1: Anti-angiogenic potential of SMYGT in vitro and in ovo. (A) HUVECs cultured on BME-pre-coated supports were exposed to different concentrations of SMYGT for 12 h. The tube length and branch numbers were determined by analyzing digitally captured images. Sulforaphane (Sulfo, 5 μM) was administered in parallel as a positive control drug. Relative tube formation was determined by comparing each group with the vehicle treated group (0 μg/mL SMYGT), and the data are presented as the means ± standard deviation (S.D.) of triplicate experiments. *P < 0.05, **P < 0.01, ***P < 0.001. (B) At ED4.5, A quarter size of thermanox coverslips containing variable amounts of SMYGT (0, 200, 400 μg/disc) were applied to the CAMs. After 2 days of incubation, 10% skimmed milk solution was injected into the CAM for observation of the inhibition zone of angiogenesis and digital images were captured. Retinoic acid (RA, 1 μg/disc) used as a control for inhibition of new vessel formation.

Mentions: The anti-angiogenic potential of SMYGT was evaluated with an in vitro tube formation assay mediated by HUVECs. In the absence of SMYGT, HUVECs constructed blood vessel-like tubes by connecting with neighboring cells (Additional file 1A, 0 μg/mL of SMYGT). However, the SMYGT treatment interrupted this intercellular connection by inhibiting tube formation (Additional file 1A, 100–400 μg/mL of SMYGT). Image analysis revealed that the degree of HUVEC-mediated angiogenesis, based on tube length and branch numbers, was decreased by SMYGT in a dose-dependent manner (Figure 1A). The inhibitory effect of 200 μg/mL of SMYGT was comparable to that of 5 μM of sulforaphane, a positive control drug, and tube formation was completely inhibited by 400 μg/mL of SMYGT.Figure 1


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)

Anti-angiogenic potential of SMYGT in vitro and in ovo. (A) HUVECs cultured on BME-pre-coated supports were exposed to different concentrations of SMYGT for 12 h. The tube length and branch numbers were determined by analyzing digitally captured images. Sulforaphane (Sulfo, 5 μM) was administered in parallel as a positive control drug. Relative tube formation was determined by comparing each group with the vehicle treated group (0 μg/mL SMYGT), and the data are presented as the means ± standard deviation (S.D.) of triplicate experiments. *P < 0.05, **P < 0.01, ***P < 0.001. (B) At ED4.5, A quarter size of thermanox coverslips containing variable amounts of SMYGT (0, 200, 400 μg/disc) were applied to the CAMs. After 2 days of incubation, 10% skimmed milk solution was injected into the CAM for observation of the inhibition zone of angiogenesis and digital images were captured. Retinoic acid (RA, 1 μg/disc) used as a control for inhibition of new vessel formation.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig1: Anti-angiogenic potential of SMYGT in vitro and in ovo. (A) HUVECs cultured on BME-pre-coated supports were exposed to different concentrations of SMYGT for 12 h. The tube length and branch numbers were determined by analyzing digitally captured images. Sulforaphane (Sulfo, 5 μM) was administered in parallel as a positive control drug. Relative tube formation was determined by comparing each group with the vehicle treated group (0 μg/mL SMYGT), and the data are presented as the means ± standard deviation (S.D.) of triplicate experiments. *P < 0.05, **P < 0.01, ***P < 0.001. (B) At ED4.5, A quarter size of thermanox coverslips containing variable amounts of SMYGT (0, 200, 400 μg/disc) were applied to the CAMs. After 2 days of incubation, 10% skimmed milk solution was injected into the CAM for observation of the inhibition zone of angiogenesis and digital images were captured. Retinoic acid (RA, 1 μg/disc) used as a control for inhibition of new vessel formation.
Mentions: The anti-angiogenic potential of SMYGT was evaluated with an in vitro tube formation assay mediated by HUVECs. In the absence of SMYGT, HUVECs constructed blood vessel-like tubes by connecting with neighboring cells (Additional file 1A, 0 μg/mL of SMYGT). However, the SMYGT treatment interrupted this intercellular connection by inhibiting tube formation (Additional file 1A, 100–400 μg/mL of SMYGT). Image analysis revealed that the degree of HUVEC-mediated angiogenesis, based on tube length and branch numbers, was decreased by SMYGT in a dose-dependent manner (Figure 1A). The inhibitory effect of 200 μg/mL of SMYGT was comparable to that of 5 μM of sulforaphane, a positive control drug, and tube formation was completely inhibited by 400 μg/mL of SMYGT.Figure 1

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