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Stimulators of mineralization limit the invasive phenotype of human osteosarcoma cells by a mechanism involving impaired invadopodia formation.

Cmoch A, Podszywalow-Bartnicka P, Palczewska M, Piwocka K, Groves P, Pikula S - PLoS ONE (2014)

Bottom Line: Here we examined the invasive potential of human osteoblast-like cells (Saos-2) and osteolytic-like (143B) OS cells treated with the stimulators of mineralization (ascorbic acid and B-glycerophosphate) and observed a significant difference in response of the tested cells to the treatment.In contrast to 143B cells, osteoblast-like cells developed a mineralization phenotype that was accompanied by a decreased proliferation rate, prolongation of the cell cycle progression and apoptosis.Herein, we show that this ability of osteolytic-like cells in vitro is limited by stimulators of mineralization.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, Poland.

ABSTRACT

Background: Osteosarcoma (OS) is a highly aggressive bone cancer affecting children and young adults. Growing evidence connects the invasive potential of OS cells with their ability to form invadopodia (structures specialized in extracellular matrix proteolysis).

Results: In this study, we tested the hypothesis that commonly used in vitro stimulators of mineralization limit the invadopodia formation in OS cells. Here we examined the invasive potential of human osteoblast-like cells (Saos-2) and osteolytic-like (143B) OS cells treated with the stimulators of mineralization (ascorbic acid and B-glycerophosphate) and observed a significant difference in response of the tested cells to the treatment. In contrast to 143B cells, osteoblast-like cells developed a mineralization phenotype that was accompanied by a decreased proliferation rate, prolongation of the cell cycle progression and apoptosis. On the other hand, stimulators of mineralization limited osteolytic-like OS cell invasiveness into collagen matrix. We are the first to evidence the ability of 143B cells to degrade extracellular matrix to be driven by invadopodia. Herein, we show that this ability of osteolytic-like cells in vitro is limited by stimulators of mineralization.

Conclusions: Our study demonstrates that mineralization competency determines the invasive potential of cancer cells. A better understanding of the molecular mechanisms by which stimulators of mineralization regulate and execute invadopodia formation would reveal novel clinical targets for treating osteosarcoma.

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Related in: MedlinePlus

Migration of osteosarcoma Saos-2 and 143B cells in the presence of ascorbic acid and β-glycerophosphate.Uniform scratches were created in confluent cell cultures, which were treated with AA/B-GP over a period of 7 days. The upper panels show representative images of wound closure in control osteosarcoma cells and cells stimulated with AA/B-GP, 12 h after scratching. Wound borders are marked with dashed lines. The lower panels represent the quantification of wound closure in % 4 h, 8 h and 12 h after injury (time 0). Time lapse imaging of wound closure was captured using a Leica AF7000 microscope Live Imaging System at 10× objective. Bars represent: open – 4 h after injury; dotted – 8 h after injury; grey stripped – 12 h after injury. The mean value for three individual experiments ± SEM is shown. Error bars indicate means ± SEM; n = 6, *p≤0.05 by Student’s t-test.
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pone-0109938-g004: Migration of osteosarcoma Saos-2 and 143B cells in the presence of ascorbic acid and β-glycerophosphate.Uniform scratches were created in confluent cell cultures, which were treated with AA/B-GP over a period of 7 days. The upper panels show representative images of wound closure in control osteosarcoma cells and cells stimulated with AA/B-GP, 12 h after scratching. Wound borders are marked with dashed lines. The lower panels represent the quantification of wound closure in % 4 h, 8 h and 12 h after injury (time 0). Time lapse imaging of wound closure was captured using a Leica AF7000 microscope Live Imaging System at 10× objective. Bars represent: open – 4 h after injury; dotted – 8 h after injury; grey stripped – 12 h after injury. The mean value for three individual experiments ± SEM is shown. Error bars indicate means ± SEM; n = 6, *p≤0.05 by Student’s t-test.

Mentions: Since cell migration is a prerequisite for tumor invasion and metastasis, we evaluated the influence of prolonged exposure of osteosarcoma cells to AA/B-GP on their migratory activities using a wound-healing assay. Fig. 4(lower panels) represents quantization of the wound closure (in %) 4, 8 and 12 h after the injury was inflicted in a 80% confluent cell monolayer, maintained in control or AA/B-GP supplemented medium for 7 days. The wound closure by Saos-2 cells after 4 h was 10.2%±2.1 in control and 2.3%±0.9 in AA/B-GP-treated cells, whilst after 12 h: 27.3%±1.9 and 17.19%±6.0, respectively. Irrespective of conditions wound closure by 143B cells was enhanced when compared to Saos-2 cells. In the case of 143B cells the wound closure 4 h post injury was 19.4%±1. 7 in control and 13.9%±1.3 in AA/B-GP-treated cells, while after 12 h it was 50.9%±2.9 and 43.3%±3.3, respectively. Therefore, regarding 143B cells, the effect of AA/B-GP on wound closure did not exceed 6% compared to control conditions. Taken together, stimulation to mineralization by the AA/B-GP treatment was accompanied by a reduced migration rate of cells of both OS cell lines.


Stimulators of mineralization limit the invasive phenotype of human osteosarcoma cells by a mechanism involving impaired invadopodia formation.

Cmoch A, Podszywalow-Bartnicka P, Palczewska M, Piwocka K, Groves P, Pikula S - PLoS ONE (2014)

Migration of osteosarcoma Saos-2 and 143B cells in the presence of ascorbic acid and β-glycerophosphate.Uniform scratches were created in confluent cell cultures, which were treated with AA/B-GP over a period of 7 days. The upper panels show representative images of wound closure in control osteosarcoma cells and cells stimulated with AA/B-GP, 12 h after scratching. Wound borders are marked with dashed lines. The lower panels represent the quantification of wound closure in % 4 h, 8 h and 12 h after injury (time 0). Time lapse imaging of wound closure was captured using a Leica AF7000 microscope Live Imaging System at 10× objective. Bars represent: open – 4 h after injury; dotted – 8 h after injury; grey stripped – 12 h after injury. The mean value for three individual experiments ± SEM is shown. Error bars indicate means ± SEM; n = 6, *p≤0.05 by Student’s t-test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196965&req=5

pone-0109938-g004: Migration of osteosarcoma Saos-2 and 143B cells in the presence of ascorbic acid and β-glycerophosphate.Uniform scratches were created in confluent cell cultures, which were treated with AA/B-GP over a period of 7 days. The upper panels show representative images of wound closure in control osteosarcoma cells and cells stimulated with AA/B-GP, 12 h after scratching. Wound borders are marked with dashed lines. The lower panels represent the quantification of wound closure in % 4 h, 8 h and 12 h after injury (time 0). Time lapse imaging of wound closure was captured using a Leica AF7000 microscope Live Imaging System at 10× objective. Bars represent: open – 4 h after injury; dotted – 8 h after injury; grey stripped – 12 h after injury. The mean value for three individual experiments ± SEM is shown. Error bars indicate means ± SEM; n = 6, *p≤0.05 by Student’s t-test.
Mentions: Since cell migration is a prerequisite for tumor invasion and metastasis, we evaluated the influence of prolonged exposure of osteosarcoma cells to AA/B-GP on their migratory activities using a wound-healing assay. Fig. 4(lower panels) represents quantization of the wound closure (in %) 4, 8 and 12 h after the injury was inflicted in a 80% confluent cell monolayer, maintained in control or AA/B-GP supplemented medium for 7 days. The wound closure by Saos-2 cells after 4 h was 10.2%±2.1 in control and 2.3%±0.9 in AA/B-GP-treated cells, whilst after 12 h: 27.3%±1.9 and 17.19%±6.0, respectively. Irrespective of conditions wound closure by 143B cells was enhanced when compared to Saos-2 cells. In the case of 143B cells the wound closure 4 h post injury was 19.4%±1. 7 in control and 13.9%±1.3 in AA/B-GP-treated cells, while after 12 h it was 50.9%±2.9 and 43.3%±3.3, respectively. Therefore, regarding 143B cells, the effect of AA/B-GP on wound closure did not exceed 6% compared to control conditions. Taken together, stimulation to mineralization by the AA/B-GP treatment was accompanied by a reduced migration rate of cells of both OS cell lines.

Bottom Line: Here we examined the invasive potential of human osteoblast-like cells (Saos-2) and osteolytic-like (143B) OS cells treated with the stimulators of mineralization (ascorbic acid and B-glycerophosphate) and observed a significant difference in response of the tested cells to the treatment.In contrast to 143B cells, osteoblast-like cells developed a mineralization phenotype that was accompanied by a decreased proliferation rate, prolongation of the cell cycle progression and apoptosis.Herein, we show that this ability of osteolytic-like cells in vitro is limited by stimulators of mineralization.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, Poland.

ABSTRACT

Background: Osteosarcoma (OS) is a highly aggressive bone cancer affecting children and young adults. Growing evidence connects the invasive potential of OS cells with their ability to form invadopodia (structures specialized in extracellular matrix proteolysis).

Results: In this study, we tested the hypothesis that commonly used in vitro stimulators of mineralization limit the invadopodia formation in OS cells. Here we examined the invasive potential of human osteoblast-like cells (Saos-2) and osteolytic-like (143B) OS cells treated with the stimulators of mineralization (ascorbic acid and B-glycerophosphate) and observed a significant difference in response of the tested cells to the treatment. In contrast to 143B cells, osteoblast-like cells developed a mineralization phenotype that was accompanied by a decreased proliferation rate, prolongation of the cell cycle progression and apoptosis. On the other hand, stimulators of mineralization limited osteolytic-like OS cell invasiveness into collagen matrix. We are the first to evidence the ability of 143B cells to degrade extracellular matrix to be driven by invadopodia. Herein, we show that this ability of osteolytic-like cells in vitro is limited by stimulators of mineralization.

Conclusions: Our study demonstrates that mineralization competency determines the invasive potential of cancer cells. A better understanding of the molecular mechanisms by which stimulators of mineralization regulate and execute invadopodia formation would reveal novel clinical targets for treating osteosarcoma.

Show MeSH
Related in: MedlinePlus