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Ganglioside GD3 induces convergence and synergism of adhesion and hepatocyte growth factor/Met signals in melanomas.

Furukawa K, Kambe M, Miyata M, Ohkawa Y, Tajima O, Furukawa K - Cancer Sci. (2013)

Bottom Line: In this study, we analyzed the effects of GD3 expression on cell signals triggered by hepatocyte growth factor (HGF)/Met interaction and by adhesion to collagen type I (CL-I).When resistance to induced apoptosis by H2O2 was examined, only GD3+ cells treated with both HGF and adhesion to CL-I showed clearly low percentages of dead cells compared with GD3- cells or GD3+ cells treated with either one of the stimulants.Cell growth measured by 5-ethynyl-2' deoxyuridine uptake also showed synergistic effects in GD3+ cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Chubu University College of Life and Health Sciences, Kasugai, Japan; Department of Biochemistry II, Nagoya University Graduate School of Medicine, Nagoya, Japan.

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Ganglioside GD3+ N1 cells showed markedly increased cell proliferation when simultaneously stimulated by hepatocyte growth factor (HGF) and collagen type I (CL-I). (A) After incubation for 16 h in serum-free medium, GD3+ cells and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with 10 ng/mL HGF and 10 μM 5-ethynyl-2 deoxyuridine (EdU). Cell proliferation was assayed with EdU uptake after incubation for 4 or 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells. (B) After incubation for 16 h in serum-free medium, GD3+ and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) and cultured with EdU (10 μM), or cultured with HGF (10 ng/mL) in non-coated 1.5 mL tubes (1.0 × 106 cells/1.0 mL) and EdU (10 μM), or plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with HGF (10 ng/mL) and EdU (10 μM). Cell proliferation was assayed with EdU uptake after incubation for 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells.
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fig08: Ganglioside GD3+ N1 cells showed markedly increased cell proliferation when simultaneously stimulated by hepatocyte growth factor (HGF) and collagen type I (CL-I). (A) After incubation for 16 h in serum-free medium, GD3+ cells and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with 10 ng/mL HGF and 10 μM 5-ethynyl-2 deoxyuridine (EdU). Cell proliferation was assayed with EdU uptake after incubation for 4 or 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells. (B) After incubation for 16 h in serum-free medium, GD3+ and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) and cultured with EdU (10 μM), or cultured with HGF (10 ng/mL) in non-coated 1.5 mL tubes (1.0 × 106 cells/1.0 mL) and EdU (10 μM), or plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with HGF (10 ng/mL) and EdU (10 μM). Cell proliferation was assayed with EdU uptake after incubation for 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells.

Mentions: To examine the implications of enhanced Erk activation in cell proliferation, we carried out the EdU uptake assay. The percentage of EdU positive cells was higher in GD3+ cells than in GD3− cells after 21 h of costimulation with HGF and adhesion to CL-I (Fig.8A). The percentage of EdU positive cells was also higher in GD3+ cells than in GD3− cells after 21 h of stimulation with adhesion to CL-I alone, whereas HGF treatment showed no significant differences in the ratios of EdU positive cells between GD3+ and GD3− cells (Fig.8B). When cells were costimulated by HGF and adhesion to CL-I, the percentage of positive cells in GD3+ cells was clearly higher than that in GD3− cells, and also than that of GD3+ cells stimulated by CL-I alone; no clear difference was found in GD3− cells between the CL-I-treated and CL-I/HGF-treated groups (Fig.8B). These results suggested that GD3 expression promotes cell proliferation with costimulation using HGF and adhesion to CL-I.


Ganglioside GD3 induces convergence and synergism of adhesion and hepatocyte growth factor/Met signals in melanomas.

Furukawa K, Kambe M, Miyata M, Ohkawa Y, Tajima O, Furukawa K - Cancer Sci. (2013)

Ganglioside GD3+ N1 cells showed markedly increased cell proliferation when simultaneously stimulated by hepatocyte growth factor (HGF) and collagen type I (CL-I). (A) After incubation for 16 h in serum-free medium, GD3+ cells and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with 10 ng/mL HGF and 10 μM 5-ethynyl-2 deoxyuridine (EdU). Cell proliferation was assayed with EdU uptake after incubation for 4 or 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells. (B) After incubation for 16 h in serum-free medium, GD3+ and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) and cultured with EdU (10 μM), or cultured with HGF (10 ng/mL) in non-coated 1.5 mL tubes (1.0 × 106 cells/1.0 mL) and EdU (10 μM), or plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with HGF (10 ng/mL) and EdU (10 μM). Cell proliferation was assayed with EdU uptake after incubation for 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4317880&req=5

fig08: Ganglioside GD3+ N1 cells showed markedly increased cell proliferation when simultaneously stimulated by hepatocyte growth factor (HGF) and collagen type I (CL-I). (A) After incubation for 16 h in serum-free medium, GD3+ cells and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with 10 ng/mL HGF and 10 μM 5-ethynyl-2 deoxyuridine (EdU). Cell proliferation was assayed with EdU uptake after incubation for 4 or 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells. (B) After incubation for 16 h in serum-free medium, GD3+ and GD3− cells were collected. These cells were suspended in FCS-free medium and rotated for 1 h at 37°C and cell suspensions were plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) and cultured with EdU (10 μM), or cultured with HGF (10 ng/mL) in non-coated 1.5 mL tubes (1.0 × 106 cells/1.0 mL) and EdU (10 μM), or plated in CL-I-coated dishes (3.0 × 105 cells/3.5-cm dish) with HGF (10 ng/mL) and EdU (10 μM). Cell proliferation was assayed with EdU uptake after incubation for 21 h. The percentages of EdU positive cells were calculated by the number of cells labeled with green fluorescence in the total number of cells.
Mentions: To examine the implications of enhanced Erk activation in cell proliferation, we carried out the EdU uptake assay. The percentage of EdU positive cells was higher in GD3+ cells than in GD3− cells after 21 h of costimulation with HGF and adhesion to CL-I (Fig.8A). The percentage of EdU positive cells was also higher in GD3+ cells than in GD3− cells after 21 h of stimulation with adhesion to CL-I alone, whereas HGF treatment showed no significant differences in the ratios of EdU positive cells between GD3+ and GD3− cells (Fig.8B). When cells were costimulated by HGF and adhesion to CL-I, the percentage of positive cells in GD3+ cells was clearly higher than that in GD3− cells, and also than that of GD3+ cells stimulated by CL-I alone; no clear difference was found in GD3− cells between the CL-I-treated and CL-I/HGF-treated groups (Fig.8B). These results suggested that GD3 expression promotes cell proliferation with costimulation using HGF and adhesion to CL-I.

Bottom Line: In this study, we analyzed the effects of GD3 expression on cell signals triggered by hepatocyte growth factor (HGF)/Met interaction and by adhesion to collagen type I (CL-I).When resistance to induced apoptosis by H2O2 was examined, only GD3+ cells treated with both HGF and adhesion to CL-I showed clearly low percentages of dead cells compared with GD3- cells or GD3+ cells treated with either one of the stimulants.Cell growth measured by 5-ethynyl-2' deoxyuridine uptake also showed synergistic effects in GD3+ cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Chubu University College of Life and Health Sciences, Kasugai, Japan; Department of Biochemistry II, Nagoya University Graduate School of Medicine, Nagoya, Japan.

Show MeSH
Related in: MedlinePlus