Journal: Breast cancer research : BCR

Article Title: The effect of versican G3 domain on local breast cancer invasiveness and bony metastasis

doi: 10.1186/bcr1751

Figure Lengend Snippet: The formation of vessel-like structures in G3-expressing cells. Culture medium in subconfluent endothelial cells was replaced with 0.5% or 1% FBS/culture medium that had been pre-incubated with G3- and vector-transfected cells for 48 h. Three (3 d) or four (4 d) days after medium change, cultures treated with G3-containing medium formed clear vessel-like structures as compared to the vector control. The scale bar depicted represents 250 μm.

Article Snippet: For these experiments, rat endothelial cells (cell line Ypen-1; CRL-222 immortalized by Adenovirus-12 SV40 hybrid virus; American Type Culture Collection, Rockville, Maryland) were used.

Techniques: Expressing, Incubation, Plasmid Preparation, Transfection, Control

Journal: Breast cancer research : BCR

Article Title: The effect of versican G3 domain on local breast cancer invasiveness and bony metastasis

doi: 10.1186/bcr1751

Figure Lengend Snippet: Effect of G3 on endothelial cells. (a) G3-containing medium enhances cell attachment and promotes greater cell viability. Culture medium in subconfluent endothelial cell cultures was replaced with 1% FBS/medium that had been preincubated with G3- or vector-transfected cells for 5 days. Two days after medium replacement, spent medium from vector-transfected cells promoted the detachment of cells from plates while spent medium from G3-transfected cells did not. (b) After cell detachment, cultures were washed, and the adherent cells were harvested and counted (mean number/field + SD). The difference in cell viability between cultures treated with medium from G3- and vector-transfected cells was significant (p < 0.01). This figure also demonstrates that G3-containing medium enhances cell proliferation and reduces cell apoptosis. (c) One day after medium change, cells treated with medium from vector-transfected cells underwent apoptosis, while cells treated with G3-containing medium grew normally. (d) DNA was prepared from cytoplasmic and nuclear fractions and subjected to agarose gel electrophoresis. DNA laddering was detected in cells treated with medium from vector-transfected cells.

Article Snippet: For these experiments, rat endothelial cells (cell line Ypen-1; CRL-222 immortalized by Adenovirus-12 SV40 hybrid virus; American Type Culture Collection, Rockville, Maryland) were used.

Techniques: Cell Attachment Assay, Plasmid Preparation, Transfection, Agarose Gel Electrophoresis, DNA Laddering

Journal: Breast cancer research : BCR

Article Title: The effect of versican G3 domain on local breast cancer invasiveness and bony metastasis

doi: 10.1186/bcr1751

Figure Lengend Snippet: Endothelial cell migration. G3-containing medium enhances endothelial cell migration 48 h following gap formation (a) . Monolayer endothelial cell cultures were wounded (0 h) by micro-tips and washed with PBS, followed by addition of culture medium pre-incubated with G3- and vector-transfected cells. Addition of G3-containing medium enhanced endothelial cell migration to the wounding areas. Endothelial cell migration is enhanced in the presence of purified G3, fibronectin (FN) and VEGF (b) . Endothelial cells were seeded on 12-well tissue culture plates. The cultures were wounded with micro-tips (before) and washed, and fibronectin, VEGF and purified G3 were added individually or in combination as indicated. In the presence of all three molecules, cells exhibited enhanced migratory capacity. Medium represents culture media in the absence of G3, VEGF, and FN.

Article Snippet: For these experiments, rat endothelial cells (cell line Ypen-1; CRL-222 immortalized by Adenovirus-12 SV40 hybrid virus; American Type Culture Collection, Rockville, Maryland) were used.

Techniques: Migration, Incubation, Plasmid Preparation, Transfection, Purification