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Hepatocyte growth factor protects endothelial cells against gamma ray irradiation-induced damage.

Hu SY, Duan HF, Li QF, Yang YF, Chen JL, Wang LS, Wang H - Acta Pharmacol. Sin. (2009)

Bottom Line: ECV304 cells derived from adult human umbilical vein endothelial cells (HUVEC) were irradiated with a single gamma ray dose of 20 Gy.Immunocytochemistry and Western blot analysis were used to detect c-Met protein expression and HGF/c-Met signal pathway.HGF significantly promoted the proliferation of ECV304 cells, and flow cytometry revealed that HGF can inhibit apoptosis of ECV304 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853,China.

ABSTRACT

Aim: To investigate the effect of HGF on proliferation, apoptosis and migratory ability of human vascular endothelial cells against gamma ray irradiation.

Methods: ECV304 cells derived from adult human umbilical vein endothelial cells (HUVEC) were irradiated with a single gamma ray dose of 20 Gy. Immunocytochemistry and Western blot analysis were used to detect c-Met protein expression and HGF/c-Met signal pathway. In the HGF-treated groups, ECV304 cells were incubated with HGF (20 or 40 ng/mL) 3 h prior to irradiation. At 48 h post-irradiation, the proliferation of ECV304 cells was measured by MTT assay, the apoptosis was assessed by flow cytometry, and the migratory ability of ECV304 cells was measured by transwell chamber assay.

Results: c-Met protein is expressed in ECV304 cells and can be activated by HGF. Gamma ray irradiation inhibits proliferation and migration of ECV304 cells in a dose-dependent manner. HGF significantly promoted the proliferation of ECV304 cells, and flow cytometry revealed that HGF can inhibit apoptosis of ECV304 cells. Transwell chamber assay also showed that HGF increases migration activity of endothelial cells.

Conclusion: HGF may afford protection to vascular endothelial cells against gamma ray irradiation-induced damage.

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Related in: MedlinePlus

c-Met protein expression and phosphorylation in ECV304 cells. (A) The ECV304 cells were immuocytochemically stained with control antibody(a) and antibody against c-Met(b) (original magnification: 100×). (B) ECV304 cells were treated with HGF at concentrations indicated for 30 min and harvested for immunoprecipitation using c-Met antibody-protein A-agarose conjugates followed by Western blot with phosphotyrosine specific antibody (top: designated as Py-c-Met). Blots were stripped and reported with antibody to c-Met (bottom).
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fig1: c-Met protein expression and phosphorylation in ECV304 cells. (A) The ECV304 cells were immuocytochemically stained with control antibody(a) and antibody against c-Met(b) (original magnification: 100×). (B) ECV304 cells were treated with HGF at concentrations indicated for 30 min and harvested for immunoprecipitation using c-Met antibody-protein A-agarose conjugates followed by Western blot with phosphotyrosine specific antibody (top: designated as Py-c-Met). Blots were stripped and reported with antibody to c-Met (bottom).

Mentions: To determine whether ECV304 cells express c-Met and whether HGF can activate signal transduction in ECV304 cells, we evaluated the expression of c-Met and phosphor-specific c-Met protein in ECV304 cells. Immunocytochemistry analysis showed ECV304 cells expressed c-Met (receptor of HGF) (Figure 1A). Further, immunoprecipitation followed by Western blot analysis, using antibodies for phosphor-tyrosine and c-Met, revealed that HGF elicited tyrosine phosphorylation of c-Met in ECV304 cells (Figure 1B).


Hepatocyte growth factor protects endothelial cells against gamma ray irradiation-induced damage.

Hu SY, Duan HF, Li QF, Yang YF, Chen JL, Wang LS, Wang H - Acta Pharmacol. Sin. (2009)

c-Met protein expression and phosphorylation in ECV304 cells. (A) The ECV304 cells were immuocytochemically stained with control antibody(a) and antibody against c-Met(b) (original magnification: 100×). (B) ECV304 cells were treated with HGF at concentrations indicated for 30 min and harvested for immunoprecipitation using c-Met antibody-protein A-agarose conjugates followed by Western blot with phosphotyrosine specific antibody (top: designated as Py-c-Met). Blots were stripped and reported with antibody to c-Met (bottom).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4007330&req=5

fig1: c-Met protein expression and phosphorylation in ECV304 cells. (A) The ECV304 cells were immuocytochemically stained with control antibody(a) and antibody against c-Met(b) (original magnification: 100×). (B) ECV304 cells were treated with HGF at concentrations indicated for 30 min and harvested for immunoprecipitation using c-Met antibody-protein A-agarose conjugates followed by Western blot with phosphotyrosine specific antibody (top: designated as Py-c-Met). Blots were stripped and reported with antibody to c-Met (bottom).
Mentions: To determine whether ECV304 cells express c-Met and whether HGF can activate signal transduction in ECV304 cells, we evaluated the expression of c-Met and phosphor-specific c-Met protein in ECV304 cells. Immunocytochemistry analysis showed ECV304 cells expressed c-Met (receptor of HGF) (Figure 1A). Further, immunoprecipitation followed by Western blot analysis, using antibodies for phosphor-tyrosine and c-Met, revealed that HGF elicited tyrosine phosphorylation of c-Met in ECV304 cells (Figure 1B).

Bottom Line: ECV304 cells derived from adult human umbilical vein endothelial cells (HUVEC) were irradiated with a single gamma ray dose of 20 Gy.Immunocytochemistry and Western blot analysis were used to detect c-Met protein expression and HGF/c-Met signal pathway.HGF significantly promoted the proliferation of ECV304 cells, and flow cytometry revealed that HGF can inhibit apoptosis of ECV304 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Chinese PLA General Hospital, Beijing 100853,China.

ABSTRACT

Aim: To investigate the effect of HGF on proliferation, apoptosis and migratory ability of human vascular endothelial cells against gamma ray irradiation.

Methods: ECV304 cells derived from adult human umbilical vein endothelial cells (HUVEC) were irradiated with a single gamma ray dose of 20 Gy. Immunocytochemistry and Western blot analysis were used to detect c-Met protein expression and HGF/c-Met signal pathway. In the HGF-treated groups, ECV304 cells were incubated with HGF (20 or 40 ng/mL) 3 h prior to irradiation. At 48 h post-irradiation, the proliferation of ECV304 cells was measured by MTT assay, the apoptosis was assessed by flow cytometry, and the migratory ability of ECV304 cells was measured by transwell chamber assay.

Results: c-Met protein is expressed in ECV304 cells and can be activated by HGF. Gamma ray irradiation inhibits proliferation and migration of ECV304 cells in a dose-dependent manner. HGF significantly promoted the proliferation of ECV304 cells, and flow cytometry revealed that HGF can inhibit apoptosis of ECV304 cells. Transwell chamber assay also showed that HGF increases migration activity of endothelial cells.

Conclusion: HGF may afford protection to vascular endothelial cells against gamma ray irradiation-induced damage.

Show MeSH
Related in: MedlinePlus