Journal: PLoS ONE

Article Title: Combined Inhibition of Epidermal Growth Factor Receptor and Cyclooxygenase-2 Leads to Greater Anti-tumor Activity of Docetaxel in Advanced Prostate Cancer

doi: 10.1371/journal.pone.0076169

Figure Lengend Snippet: Cells grown in 1% FBS were exposed to 0.01 μmol/L docetaxel (D), 20 μmol/L gefitinib (G) or100 μmol/LNS-398 (N), alone or in combination, or with DMSO as control for 24 h. NF-KB, MMP-9 and VEGF mRNA (A and B) and protein levels (C and D) were measured as described in Materials and Methods. Values are reported as the mean±SD of three independent experiments.

Article Snippet: Antibodies against human VEGF and MMP-9 were obtained from R&D System and Bioworld, respectively.

Techniques: Control

Journal: Cells

Article Title: Effects of Human Deciduous Dental Pulp-Derived Mesenchymal Stem Cell-Derived Conditioned Medium on the Metabolism of HUVECs, Osteoblasts, and BMSCs

doi: 10.3390/cells11203222

Figure Lengend Snippet: Effect of VEGF-A in SHED-CM on the proliferative ability of HUVECs. The cells in the SHED-CM group exhibited significantly higher proliferative ability than those in the SHED-CM + anti-VEGF-A neutralizing antibody-supplemented (hereafter termed SHED-CM + anti-VEGF-A), VEGF-A, and control groups. Cells in the SHED-CM + anti-VEGF-A and VEGF-A groups showed significantly higher proliferative ability than those in the control group. ( n = 12, ** p < 0.01, * p < 0.05). SHED-CM, human deciduous dental pulp-derived mesenchymal stem cell-derived culture supernatant; VEGF-A, vascular endothelial growth factor-A; HUVECs, human umbilical vein endothelial cells.

Article Snippet: HUVECs were treated with SHED-CM in the presence or absence of 10 μg/mL anti-VEGF-A antibody (MAB293; R&D Systems, Minneapolis, MN, USA) in serum-free EBM-2 medium.

Techniques: Control, Derivative Assay

Journal: Cells

Article Title: Effects of Human Deciduous Dental Pulp-Derived Mesenchymal Stem Cell-Derived Conditioned Medium on the Metabolism of HUVECs, Osteoblasts, and BMSCs

doi: 10.3390/cells11203222

Figure Lengend Snippet: Effects of VEGF-A in SHED-CM on HUVEC lumen assembly. ( A ) Luminal view of HUVECs in the four groups. Lumen formation was observed more frequently in the SHED-CM- and VEGF-A-supplemented groups. Scale bar = 300 μm. ( B ) Vascular bifurcation in the four groups. ( C ) Vessel length in the four groups ( n = 4, ** p < 0.01). Black lines indicate vessel length; black arrows indicate joints. SHED-CM, human deciduous dental pulp-derived mesenchymal stem cell-derived culture supernatant; VEGF-A, vascular endothelial growth factor-A; HUVECs, human umbilical vein endothelial cells.

Article Snippet: HUVECs were treated with SHED-CM in the presence or absence of 10 μg/mL anti-VEGF-A antibody (MAB293; R&D Systems, Minneapolis, MN, USA) in serum-free EBM-2 medium.

Techniques: Derivative Assay

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Vascular endothelial growth factor confers endothelial resistance to apoptosis through poly(ADP-ribose) polymerase.

doi: 10.1111/j.1538-7836.2011.04368.x

Figure Lengend Snippet: Fig. 2. Vascular endothelial growth factor (VEGF) time-dependently increases resistance to apoptosis. (A) Preincubation of human umbilical vein endothelial cells (HUVECs) grown on vitronectin with 100 pg mL)1 to 100 ng mL)1 (0.1–100 ng mL)1) VEGF-A(165) for 24 h significantly reduced apoptosis induced by 5 lg mL)1 cRGDfV (RGD) by an average of 64.8% ± 4.0% (*P < 0.05) as compared with incubation with cRGDfV only, as demonstrated by flow cytometry. Apoptotic cells were considered annexin V+/PI) cells. Mean ± standard deviation (SD) of three experiments in duplicate. (B) Longer-term preincubation of HUVECs with 100 pg mL)1 to 100 ng mL)1 (0.1–100 ng mL)1) VEGF-A(165) at 48-h intervals for 6 days further reduced cRGDfV-induced apoptosis significantly, by an average of 90.3% ± 3.3% (*P < 0.05). Results were reproduced in hCMEC/D3 and EA.hy.926 cells. Mean ± SD of four experiments in duplicate. (C) Preincubation of HUVECs with VEGF-A(165) for 6 days (48-h intervals) reduced the number of non-vital cells in trypan blue exclusion assays as compared with cRGDfV only, and (D) reduced apoptotic chromatin condensation and fragmentation as analyzed by diamidino-2-phenylindole (DAPI)-stained nuclear morphology, corroborating the results from flow cytometry. Mean ± SD of three experiments in duplicate. The small panel in (D) shows the DAPI staining of a representative experiment: HUVECs incubated with 10 ng mL)1

Article Snippet: They were then incubated with 5% normal serum/PBS, and then with goat-anti-human PARP (1 : 20; R&D Systems), goat anti-human VEGF (1 : 50; R&D Systems), mouse anti-human VEGFR-2 (clone Y-23, 1 lg mL)1; Santa Cruz), rabbit-antihuman NP-1 (5 lg mL)1; Invitrogen) and rabbit-anti-human von Willebrand factor (VWF) (1 : 200; Dako) antibodies for 2 or 16 h at 4 C, and subsequently with biotinylated secondary antibodies (streptavidin–alkaline phosphatase conjugate, Fast Red, Mayer s hemalum counterstaining [all Sigma-Aldrich]), and covered with glycerol gelatine.

Techniques: Incubation, Cytometry, Standard Deviation, Staining

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Vascular endothelial growth factor confers endothelial resistance to apoptosis through poly(ADP-ribose) polymerase.

doi: 10.1111/j.1538-7836.2011.04368.x

Figure Lengend Snippet: Fig. 5. Vascular endothelial growth factor (VEGF) induces poly(ADP-ribose) polymerase-1 (PARP) expression through VEGF receptor 2 (VEGFR-2), Akt and c-Jun N-terminal kinase (JNK) activation. (A) Intracellular signaling was analyzed by short-term exposure of human umbilicalvein endothelial cells (HUVECs)to 10and 100 ng mL)1 VEGF-A(165) for15–120 min. Kinase phosphorylationwasdemonstratedbyautomated densitometryof Western blots; phospho-kinases were correlated with the total amount of the respective kinase expressed as ratio of relative optical density (ROD). VEGF significantly induced time-dependent and dose-dependent phosphorylation of JNK and Akt as compared with negative controls (P < 0.05), whereas p38 mitogen- activated protein kinase and extracellular signal-related kinase 1/2 (p42/44) remained uninfluenced. Mean (%) ± standard deviation (SD) of four exper- iments in duplicate. (B) VEGF receptor signal transduction responsible for PARP regulation was analyzed by incubation of HUVECs with 10 ng mL)1

Article Snippet: They were then incubated with 5% normal serum/PBS, and then with goat-anti-human PARP (1 : 20; R&D Systems), goat anti-human VEGF (1 : 50; R&D Systems), mouse anti-human VEGFR-2 (clone Y-23, 1 lg mL)1; Santa Cruz), rabbit-antihuman NP-1 (5 lg mL)1; Invitrogen) and rabbit-anti-human von Willebrand factor (VWF) (1 : 200; Dako) antibodies for 2 or 16 h at 4 C, and subsequently with biotinylated secondary antibodies (streptavidin–alkaline phosphatase conjugate, Fast Red, Mayer s hemalum counterstaining [all Sigma-Aldrich]), and covered with glycerol gelatine.

Techniques: Expressing, Activation Assay, Western Blot, Phospho-proteomics, Standard Deviation, Transduction, Incubation

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Vascular endothelial growth factor confers endothelial resistance to apoptosis through poly(ADP-ribose) polymerase.

doi: 10.1111/j.1538-7836.2011.04368.x

Figure Lengend Snippet: Fig. 6. Poly(ADP-ribose) polymerase-1 (PARP) expression in an organ culture model. Vascular endothelial growth factor (VEGF) increased endothelial PARP expression in a human vascular organ culture model implemented in internal mammary artery (IMA) segments from nine pa- tients in duplicate or triplicate, as demonstrated by immunohistochemistry. The panels show a representative IMA specimen preculture (Pre), control incubation(Co)andincubationwith10 ng mL)1VEGF-A(165)for6 days. Endothelial PARP expression is indicated by arrowheads. Magnification: · 400. Scale bar: 100 lm. End, endothelium; Int, intima; Med, media.

Article Snippet: They were then incubated with 5% normal serum/PBS, and then with goat-anti-human PARP (1 : 20; R&D Systems), goat anti-human VEGF (1 : 50; R&D Systems), mouse anti-human VEGFR-2 (clone Y-23, 1 lg mL)1; Santa Cruz), rabbit-antihuman NP-1 (5 lg mL)1; Invitrogen) and rabbit-anti-human von Willebrand factor (VWF) (1 : 200; Dako) antibodies for 2 or 16 h at 4 C, and subsequently with biotinylated secondary antibodies (streptavidin–alkaline phosphatase conjugate, Fast Red, Mayer s hemalum counterstaining [all Sigma-Aldrich]), and covered with glycerol gelatine.

Techniques: Expressing, Organ Culture, Immunohistochemistry, Control, Incubation

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Vascular endothelial growth factor confers endothelial resistance to apoptosis through poly(ADP-ribose) polymerase.

doi: 10.1111/j.1538-7836.2011.04368.x

Figure Lengend Snippet: Fig. 7. Poly(ADP-ribose) polymerase-1 (PARP) expression in human atherosclerotic and normal arteries. (A) PARP expression was compared in 24 atherosclerotic (11 carotid, 13 femoral/iliac specimens) vs. 10 normal arteries by immunohistochemistry (see also Table 1). The panels show representative atherosclerotic (femoral) and normal (internal mammary artery) specimens demonstrating PARP and von Willebrand factor (VWF) immunostaining (arrowheads). Magnification: · 200. In atherosclerosis, endothelial PARP immunostaining was increased as compared with nor- mal arteries. (B) Local vascular endothelial growth factor (VEGF) expression was found in atherosclerotic arteries (magnification: · 100). Strong VEGF immunostaining in the lamina media (black arrowheads) and thrombocyte aggregates (open arrowhead) is shown in a representa- tive atherosclerotic specimen. In (A) and (B), immunostaining was visu- alized by alkaline phosphatase conjugate, Fast Red staining, and hemalum counterstaining, and documented with the use of a Zeiss Axio Scope microscope, AXIOVISION software (Zeiss), and IMAGE J (freeware). Vas- cular structures are indicated as follows: End, endothelium; Int, intima; Med, media; Neo, atherosclerotic neointima. The scale bars in (A) and (B) represent 100 lm. The results underline the significance of a local vascular VEGF effect for increased endothelial PARP expression in human arteries and in the vascular culture model.

Article Snippet: They were then incubated with 5% normal serum/PBS, and then with goat-anti-human PARP (1 : 20; R&D Systems), goat anti-human VEGF (1 : 50; R&D Systems), mouse anti-human VEGFR-2 (clone Y-23, 1 lg mL)1; Santa Cruz), rabbit-antihuman NP-1 (5 lg mL)1; Invitrogen) and rabbit-anti-human von Willebrand factor (VWF) (1 : 200; Dako) antibodies for 2 or 16 h at 4 C, and subsequently with biotinylated secondary antibodies (streptavidin–alkaline phosphatase conjugate, Fast Red, Mayer s hemalum counterstaining [all Sigma-Aldrich]), and covered with glycerol gelatine.

Techniques: Expressing, Immunohistochemistry, Immunostaining, Staining, Microscopy, Software

Journal: European radiology

Article Title: Magnetic resonance imaging for monitoring the effects of thalidomide on experimental human breast cancers

doi: 10.1007/s00330-008-1111-x

Figure Lengend Snippet: Representative immunohistochemically stained human breast cancer sections (MDA-MB-435) showing leakage of macromolecular contrast medium (streptavidin-biotin reaction), vascular richness (lectin and RECA-1 antibody), and VEGF in tumor cells. Thalidomide treatment reduces the extravasation of albumin-(Gd-DTPA)27-(biotin)11 without reducing the abundance of tumor blood vessels. a Tumor section from the saline-control group administered albumin-(Gd-DTPA)27-(biotin)11. The strong red signal indicates extravascular 1-h accumulation of biotin-labeled contrast medium surrounding the yellow-green tumor microvessels. b After 7 days of treatment with thalidomide, the density of red-fluorescent, biotin-labeled contrast agent extravasated over 1 h is strongly reduced compared with a, indicating a reduction in leakage and extravascular accumulation of the macromolecules. Confocal microscopic images of tumor vessels in MDA-MB-435 tumors after treatment with saline (c, d) or thalidomide (e, f) for 7 days show no noticeable change in the area density of perfused blood vessels (green lectin-stained) or total blood vessels (red, RECA-1 stained). No difference is observable (c–f) between saline-control and thalidomide-treated groups with regard to tumor vascularity. (g, h). Representative MDA-MB-435 tumor sections after immunohistochemical staining for human VEGF after a 7-day, three-injection treatment protocol with saline (g) or thalidomide (h). No difference in amount VEGF immunoreactivity was detected in the two groups. Scale bar 115 μm in c–f and 120 μm in a, b

Article Snippet: VEGF immunoreactivity of the human tumor cells was detected using goat polyclonal antibody anti-human VEGF (1:500, R&D Systems, Minneapolis, Minn.).

Techniques: Staining, Saline, Control, Labeling, Immunohistochemical staining, Injection

Journal: European radiology

Article Title: Magnetic resonance imaging for monitoring the effects of thalidomide on experimental human breast cancers

doi: 10.1007/s00330-008-1111-x

Figure Lengend Snippet: Measurement of RECA-1 and VEGF area density in tumors treated with thalidomide or saline control. Values obtained after 7 days of therapy

Article Snippet: VEGF immunoreactivity of the human tumor cells was detected using goat polyclonal antibody anti-human VEGF (1:500, R&D Systems, Minneapolis, Minn.).

Techniques: Saline, Control