The vascular endothelial growth factor (VEGF) family of soluble protein growth factors consists of key mediators of angiogenesis and lymphangiogenesis in the context of tumor biology. The members of the family, VEGF-A (also known as VEGF), VEGF-B, VEGF-C, VEGF-D, and placenta growth factor (PlGF), play important roles in vascular biology in both normal physiology and pathology. The generation of a humanized neutralizing antibody to VEGF-A (bevacizumab, also known as Avastin) and the demonstration of its benefit in numerous human cancers have confirmed the merit of an anti-angiogenesis approach to cancer treatment and have validated the VEGF-A signaling pathway as a therapeutic target. Other members of the VEGF family are now being targeted, and their relevance to human cancer and the development of resistance to anti-VEGF-A treatment are being evaluated in the clinic. Here, we discuss the potential of targeting VEGF family members in the diagnosis and treatment of cancer.
Angiogenesis Inhibitors ; therapeutic use ; Animals ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Bevacizumab ; Humans ; Lymphangiogenesis ; Neoplasms ; drug therapy ; metabolism ; Neovascularization, Pathologic ; metabolism ; Placenta Growth Factor ; Pregnancy Proteins ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Signal Transduction ; drug effects ; Vascular Endothelial Growth Factor A ; classification ; metabolism ; Vascular Endothelial Growth Factor B ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVETo investigate the effects of bortezomib on the migration of endothelial cells and the expression of angiogenesis-related molecules, and explore the mechanism of its antiproliferation of tumor cells.

METHODSCell count kit CCK-8 was used to detect the relative proliferation activity of cells after treated by bortezomib at different concentrations for 12 h and 24 h, respectively. Transwell model was uesd to detect the migration rate of cells. Expression levels of VEGF and Annexin A2 genes were determined by real-time quantitative PCR. Annexin A2 protein was validated by Western blot.

RESULTSAfter treated with bortezomib at concentrations of 2.5, 5.0 and 10 nmol/L for 12h, respectively, the HMEC-1 cell proliferation activity was 1.004 ± 0.002, 0.793 ± 0.021 and 0.874 ± 0.062, respectively, being no statistical difference from that of control group (1.000) P < 0.05); while the migration rates of them were 0.697 ± 0.060, 0.597 ± 0.090 and 0.874 ± 0.062, respectively, being significantly lower than that of control group (1.000) (P < 0.05) and so did for the expression of VEGF and Annexin A2 genes. After treated with 5 nmol/L bortezomib for 12 h, the Annexin A2 and VEGF gene relative expression level of HMEC-1 cells was 0.540 ± 0.001 and 0.793 ± 0.153, respectively, being of statistical difference from that of control group (1.000) P < 0.05). The conspicuous downregulation of Annexin A2 protein was also confirmed by Western Blot.

CONCLUSIONSBortezomib can inhibit migration of endothelial cell HMEC-1 by downregulating the expression of VEGF and Annexin A2, displaying a new mechanism of bortezomib for inhibition of tumor proliferation.

Annexin A2 ; metabolism ; Bortezomib ; Cell Proliferation ; drug effects ; Endothelial Cells ; metabolism ; Humans ; RNA, Messenger ; genetics ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo explore the effect of tacrolimus on murine hair follicle cycle.

METHODHematoxylin-eosin dyeing and reverse transcription-polymerase chain raction techniques were used.

RESULTSFive days after depilation, the hair follicles in both the tacrolimus group and the minoxidil group was in anagen V, while that in the vaseline group was in anagen III. vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were detected in back skin in both the tacrolimus group and the minoxidil group, but not in the vaseline group.

CONCLUSIONTacrolimus can promote the growth of hair by stimulating the hair follicle to enter anagen V in mice, which may be explained by the effects of VEGF and HGF.

Animals ; Hair Follicle ; drug effects ; physiology ; Hepatocyte Growth Factor ; metabolism ; Mice ; Mice, Inbred C57BL ; Minoxidil ; pharmacology ; Skin ; drug effects ; metabolism ; Tacrolimus ; pharmacology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effect of vasoactive intestinal peptide (VIP) on angiogenesis after focal cerebral ischemia.

METHODSFocal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 120 min in adult SD rats with intracerebroventricular VIP administration at the beginning of reperfusion. Immunohistochemistry was performed to assay BrdU immunoreactive endothelial cells, expressions of VEGF, flt-1 and flk-1 in the ischemic zone, and the protein expressions of vascular endothelial growth factor (VEGF) in the brain was measured using Western blotting.

RESULTSImmunohistochemical staining revealed significantly increased BrdU immunoreactive endothelial cells on the margins of the ischemic lesion in rats treated with VIP as compared with that in the control rats (P<0.05). VIP significantly increased the number of VEGF immunoreactive cells and flt-1- and flk-1-positive endothelial cells in comparison with the control group (P<0.01). Western blotting showed that VIP treatment resulted in significantly increased VEGF protein level in the ipsilateral hemisphere (P<0.05).

CONCLUSIONSVIP enhances angiogenesis in the ischemic brain by increasing the expressions of VEGF in the brain tissue and its receptors flt-1 and flk-1 in the endothelial cells.

Animals ; Brain ; drug effects ; metabolism ; Brain Ischemia ; immunology ; metabolism ; pathology ; physiopathology ; Endothelial Cells ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Male ; Neovascularization, Physiologic ; drug effects ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism ; Vasoactive Intestinal Peptide ; pharmacology

OBJECTIVETo study differentiation of human bone marrow-derived mesenchymal stem cells (BDMSC) into blood vessel endothelial cells for ideal cell origin of complex organ tissue engineering vascularization and injured tissue repairing by cell transplantation.

METHODAfter different days of induction with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in 3D fibrin-gels and matrigel, BDMSC and angiogenesis were determined by the utilization of morphological observation, tissue section and CD34, CD31, vascular endothelial growth factor receptor-1 (VEGFR-1, Flt-1), VEGFR-2 (Flk-1), and vWF that were special for blood vessel endothelial cells.

RESULTAfter 3D-cultured and induced with VEGF and bFGF in vitro in fibrin-gels and matrigel for 3-21 days, BDMSC expressed CD34, CD31, Flt-1, Flk-1, and vWF came into vessel-like configuration.

CONCLUSIONVEGF, bFGF as well as Flt-1 and Flk-1, expressed by BDMSC, may form a feasible microenviroment after induction and play an important role during processes of blood vessel endothelial cell differentiation and vessel-like configuration forming of BDMSC. Mesenchymal stem cells may be applied to tissue engineering vascularization and injured tissue repairing by cell transplantation.

Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Endothelial Cells ; cytology ; drug effects ; Fibrin ; Fibroblast Growth Factor 2 ; pharmacology ; Gels ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Vascular Endothelial Growth Factor A ; pharmacology ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism ; von Willebrand Factor ; metabolism

OBJECTIVETo study the effect of insulin in different concentrations on secretion function of growth factors of adipose-derived stem cells (ADSCs).

METHODSADSCs were isolated from human abdominal adipose tissue and cultured. The immunophenotype and adipose induced-differentiation were identified, and the third generation cells were collected. The collected cells were assigned to 1 x 10(-8), 1 x 10(-7), 1 x 10(-6) mol/L insulin groups according to the concentration of added insulin. When cells grew into 70% confluence in conventional medium, ADSCs were cultured further in serum-free DMEM containing insulin in different concentrations for 3 days. ADSCs cultured in medium without insulin were used as control group. Secretion amount of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) of ADSCs were determined by enzyme-linked immunosorbent assay. The effects of the supernatant fluid of ADSCs' nutrient solution on the proliferation and collagen synthesis of the cultured fibroblast were detected by MTT chromatometry and hydroxyproline chromatometry.

RESULTSThe secretion amounts of VEGF and HGF of ADSCs in 1 x 10(-8) and 1 x 10(-7) mol/L insulin groups [(471 +/- 41, 762 +/- 66 ng/L), (643 +/- 64, 930 +/- 67 ng/L), respectively] were significantly higher as compared with those in control group (286 +/- 47, 577 +/- 84 ng/L) (P < 0.05 or P < 0.01). No change occurred in the secretion amount of VEGF and HGF of ADSCs in 1 x l0(-6) mol/L insulin group (P > 0.05). The supernatant fluid of ADSCs' nutrient medium of 1 x 10(-8), 1 x 10(-7) mol/L insulin groups showed obvious stimulative effect on the proliferation and collagen synthesis of fibroblasts, and it was most obvious in the 1 x 10(-7) mol/L group (P < 0.05 or P < 0.01).

CONCLUSIONSInsulin in the concentrations of 1 x 10(-8) and 1 x 10(-7) mol/L can notably promote ADSCs' function of secreting VEGF and HGF.

Adipocytes ; cytology ; drug effects ; secretion ; Cells, Cultured ; Fibroblasts ; cytology ; Hepatocyte Growth Factor ; metabolism ; Humans ; Insulin ; pharmacology ; Stem Cells ; cytology ; drug effects ; secretion ; Vascular Endothelial Growth Factor A ; metabolism

BACKGROUNDCelastrol is a major active component of Tripterygium wilfordii named "Thunder God Vine", which is widely used to treat rheumatoid arthritis in China. The present study aims to demonstrate that celastrol has potent anticancer activity against glioma in vitro and in vivo.

METHODSProliferation, migration, and tube formation of ECV-304 cells were determined by MTT and matrigel assays. The antiangiogenesis effect of celastrol was assessed by the chick chorioallantoic membrane assay and the in vivo matrigel plug assay. Tumor microvessels (MVD) were determined immunohistochemically with anti-CD34 antibody. Vascular endothelial growth factor (VEGF) expression was defined as positive if distinct staining of the cytoplasm was observed in at least 10% of tumor cells at the deepest invasive site, central portion and superficial part of the tumor. MVD was estimated by averaging the counts of three times at a x 200 field in the most vascularized area of the deepest invasive site.

RESULTSCelastrol purified from T. wilfordii inhibited the proliferation of vascular endothelial cells (ECV-304) with an IC50 value of 1.33 microg/ml. Celastrol, at the concentration of 0.2 microg/ml, significantly inhibited cell migration and tube formation. Celastrol inhibited angiogenesis in a dose-dependent manner both in vitro and in vivo. Subcutaneous administration of celastrol 5 days a week for 4 consecutive weeks significantly reduced tumor volume in a dose-dependent manner in the SHG-44 xenograft model. Celastrol at each different dose level lowered the density of MVD significantly in tumor bearing nude mice compared to the control group. Immunohistochemistry experiments further revealed that celastrol also decreased the level of VEGFR-1 and VEGFR-2 expression, but not the level of VEGF expression.

CONCLUSIONSCelastrol elicits antiangiogenic effects in vitro and in vivo, and could be of potential use in the treatment of malignant cancers such as glioblastoma.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Chick Embryo ; Chickens ; Chorioallantoic Membrane ; drug effects ; Female ; Glioma ; drug therapy ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; drug effects ; Triterpenes ; pharmacology ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo study the effect of adenovirus vector-mediated siRNA targeting vascular endothelial growth factor(VEGF) on apoptosis and the expression of survivin in K562 cells.

METHODSK562 cells were infected with recombinant adenovirus Ad5-VEGFsi for 72 hours as experimental group (K562/Ad5-VEGFsi), and empty vector group (K562/Ad5) and blank control group (K562) as controls. VEGF mRNA and survivin mRNA expression were determined by RT-PCR. The protein levels of VEGF and survivin were measured by ELISA and Western blot, respectively. The apoptosis of K562 cells was detected by flow cytometry.

RESULTSThe levels of VEGF and survivin mRNA expression in experimental group cells were significantly decreased (P < 0.01). The protein concentration of VEGF in experimental group supernatant was (1121 +/- 15) pg/ml, being lower than that in empty vector group \[(1290 +/- 28) pg/ml\] and black control group \[(1303 +/- 28) pg/ml\] (P < 0.01), and the level of survivin protein in experimental group (0.26 +/- 0.11) was significantly reduced compared with that in blank control group (0.74 +/- 0.10) (P < 0.01). The apoptosis rate of K562/Ad5-VEGFsi cells (16.45 +/- 0.14)% was higher than those of K562/Ad5 cells (3.54 +/- 0.17)% and K562 cells (2.56 +/- 0.20)% (P < 0.01).

CONCLUSIONSVEGF can up-regulate the expression of survivin. After inhibition of VEGF by RNAi, the expression of survivin is decreased subsequently and the rates of cell apoptosis are increased.

Apoptosis ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; K562 Cells ; RNA, Small Interfering ; genetics ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo conduct a preliminary study on the effect of curcumol in promoting angiogenesis activity and its mechanism in zebrafishes, in order to provide basis for clinical prescription.

METHODZebrafishes biological model was established to, observe curcumol's effect on embryo blood vessel growth, blood vessel regeneration of adult fishes after tail-cutting and tissue regeneration of fish fries after tail-cutting. The relative fluorescence quantitative PCR method was adopted to determine the gene expression of vascular endothelial growth factor (VEGFA) and receptor VEGFR2 of fish fries after tail-cutting.

RESULTCurcumol contributed to angiogenesis of intersegmental blood vessels in zebrafishes embryos and speed up regeneration of blood vessels in adult fishes after tail-cutting. Furthermore, curcumol can increase the gene expression of VEGFA and VEGFR2 in fish fries.

CONCLUSIONCurcumol can promote angiogenesis in zebrafishes, and enhance the gene expression of VEGFA and VEGFR2 in fish fries after tail-cutting and speed up the regeneration of their tails.

Animals ; Embryo, Nonmammalian ; blood supply ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Neovascularization, Physiologic ; drug effects ; Sesquiterpenes ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; Zebrafish ; embryology ; genetics ; physiology

OBJECTIVETo investigate the effect of Hirudin on random skin flap survival in rats.

METHODS24 SD rats were randomly divided into control group and experimental group. The "McFarlane flap (3 cm x 9 cm)" rat models were established on the rat dorsum. 3 ml Hirudin (30 ATU) was injected into the flap in the experimental group, while 3 ml saline in the control group. The injection was performed for 7 days. The flap survival area in the two groups was measured. The tissue samples were taken from proximal (I), middle (II) and distal (III) portions of flaps for histologic study. The VEGF and bFGF expression was also detected with immunohistochemistry method.

RESULTS7 days after operation, the flap survival rate was (69.52 +/- 3.23)% in the experimental group, while (50.36 +/- 2.37)% in control group, showing a significant difference between the two groups (P < 0. 01). In the middle portion, tissue edema and infiltration of neutrophils in experimental group was markedly slighter than that in control group. The VEGF and bFGF expression and neovascularization was enhanced markedly in experimental group.

CONCLUSIONSHirudin can increase the survival of random pattern skin flaps. It may increase the VEGF, bFGF expression through a series of complex regulatory pathway. Then flap neovascularization is promoted and the flap blood supply is increased.

Animals ; Fibroblast Growth Factor 2 ; metabolism ; Graft Survival ; drug effects ; Hirudins ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; Surgical Flaps ; Vascular Endothelial Growth Factor A ; metabolism