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Syntenin controls migration, growth, proliferation, and cell cycle progression in cancer cells.

Kashyap R, Roucourt B, Lembo F, Fares J, Carcavilla AM, Restouin A, Zimmermann P, Ghossoub R - Front Pharmacol (2015)

Bottom Line: In human adulthood, syntenin gain-of-function is increasingly associated with various cancers and poor prognosis.We observed decreased migration, growth, and proliferation of the mouse melanoma cell line B16F10, the human colon cancer cell line HT29 and the human breast cancer cell line MCF7.We further documented that syntenin controls the presence of active β1 integrin at the cell membrane and G1/S cell cycle transition as well as the expression levels of CDK4, Cyclin D2, and Retinoblastoma proteins.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Signal Integration in Cell Fate Decision, Department of Human Genetics, KU Leuven Leuven, Belgium ; Centre de Recherche en Cancérologie de Marseille, Aix-Marseille Université Marseille, France ; Inserm U1068, Institut Paoli-Calmettes Marseille, France ; Centre National de la Recherche Scientifique, UMR7258 Marseille, France.

ABSTRACT
The scaffold protein syntenin abounds during fetal life where it is important for developmental movements. In human adulthood, syntenin gain-of-function is increasingly associated with various cancers and poor prognosis. Depending on the cancer model analyzed, syntenin affects various signaling pathways. We previously have shown that syntenin allows syndecan heparan sulfate proteoglycans to escape degradation. This indicates that syntenin has the potential to support sustained signaling of a plethora of growth factors and adhesion molecules. Here, we aim to clarify the impact of syntenin loss-of-function on cancer cell migration, growth, and proliferation, using cells from various cancer types and syntenin shRNA and siRNA silencing approaches. We observed decreased migration, growth, and proliferation of the mouse melanoma cell line B16F10, the human colon cancer cell line HT29 and the human breast cancer cell line MCF7. We further documented that syntenin controls the presence of active β1 integrin at the cell membrane and G1/S cell cycle transition as well as the expression levels of CDK4, Cyclin D2, and Retinoblastoma proteins. These data confirm that syntenin supports the migration and growth of tumor cells, independently of their origin, and further highlight the attractiveness of syntenin as potential therapeutic target.

No MeSH data available.


Related in: MedlinePlus

Syntenin loss-of-function leads to reduced growth in different cancer cell models. (A) Representative images of colony formation assays with the different cell models. (B) Bar graphs indicate the number of colonies formed relative to colony numbers formed by empty vector-transduced cells (taken as 100%). n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test). (C) Representative images of soft agar colony formation assays with the different cell models. (D) Bar graphs indicate luciferase activities expressed in relative light units (RLUs), relative to activities measured in empty vector transduced cells (taken as 100%). Note that syntenin-depleted cells grow more slowly than the control cells. n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test).
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Figure 3: Syntenin loss-of-function leads to reduced growth in different cancer cell models. (A) Representative images of colony formation assays with the different cell models. (B) Bar graphs indicate the number of colonies formed relative to colony numbers formed by empty vector-transduced cells (taken as 100%). n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test). (C) Representative images of soft agar colony formation assays with the different cell models. (D) Bar graphs indicate luciferase activities expressed in relative light units (RLUs), relative to activities measured in empty vector transduced cells (taken as 100%). Note that syntenin-depleted cells grow more slowly than the control cells. n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test).

Mentions: The role of syntenin in cell growth was investigated using in vitro plate colony formation assays (Figures 3A,B) and soft agar colony formation assays (Figures 3C,D). The number of colonies was significantly decreased in syntenin-depleted B16F10, HT29, and MCF7 cells compared to control cells, i.e., cells expressing empty vectors. In MCF7 cells, two different shRNA induced a drastic reduction of plate colony formation, decreasing, respectively, by 85 and 88% compared to control. In B16F10 and HT29 cells, the reduction of colony numbers was less drastic but still significant, with a decrease by 30% in B16F10 and by 36 or 38% in HT29 cells depending on the shRNA (Figures 3A,B). We also performed soft agar colony formation assays to test cellular abilities to form colonies in three dimensions because this assay is considered as the most stringent test for the detection of anchorage-independent tumor cell growth (Horibata et al., 2015). shRNA targeting syntenin induced a 49% decrease in the growth of B16F10 cells, a 20 or 33% decrease in the growth of HT29 cells and a 44 or 46% decrease in the growth of MCF7 cells (respective values for the two different shRNA) (Figures 3C,D). Control shRNA, used as control, did not significantly affect colony formation, neither in plate nor in soft agar assays, when compared to empty vector transduced B16F10 and HT29 cells. Surprisingly, control shRNA significantly affected colony formation of MCF7 cells in both assays. The reason for this specific effect in MCF7 cells is unknown. While a priori this observation does not change the conclusion that syntenin depletion affects the growth of MCF7 cells, it indicates that from a quantitative point of view, data should be interpreted with caution.


Syntenin controls migration, growth, proliferation, and cell cycle progression in cancer cells.

Kashyap R, Roucourt B, Lembo F, Fares J, Carcavilla AM, Restouin A, Zimmermann P, Ghossoub R - Front Pharmacol (2015)

Syntenin loss-of-function leads to reduced growth in different cancer cell models. (A) Representative images of colony formation assays with the different cell models. (B) Bar graphs indicate the number of colonies formed relative to colony numbers formed by empty vector-transduced cells (taken as 100%). n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test). (C) Representative images of soft agar colony formation assays with the different cell models. (D) Bar graphs indicate luciferase activities expressed in relative light units (RLUs), relative to activities measured in empty vector transduced cells (taken as 100%). Note that syntenin-depleted cells grow more slowly than the control cells. n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test).
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Figure 3: Syntenin loss-of-function leads to reduced growth in different cancer cell models. (A) Representative images of colony formation assays with the different cell models. (B) Bar graphs indicate the number of colonies formed relative to colony numbers formed by empty vector-transduced cells (taken as 100%). n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test). (C) Representative images of soft agar colony formation assays with the different cell models. (D) Bar graphs indicate luciferase activities expressed in relative light units (RLUs), relative to activities measured in empty vector transduced cells (taken as 100%). Note that syntenin-depleted cells grow more slowly than the control cells. n = 3, bars represent mean value ± SD, n.s., non-significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student’s t-test).
Mentions: The role of syntenin in cell growth was investigated using in vitro plate colony formation assays (Figures 3A,B) and soft agar colony formation assays (Figures 3C,D). The number of colonies was significantly decreased in syntenin-depleted B16F10, HT29, and MCF7 cells compared to control cells, i.e., cells expressing empty vectors. In MCF7 cells, two different shRNA induced a drastic reduction of plate colony formation, decreasing, respectively, by 85 and 88% compared to control. In B16F10 and HT29 cells, the reduction of colony numbers was less drastic but still significant, with a decrease by 30% in B16F10 and by 36 or 38% in HT29 cells depending on the shRNA (Figures 3A,B). We also performed soft agar colony formation assays to test cellular abilities to form colonies in three dimensions because this assay is considered as the most stringent test for the detection of anchorage-independent tumor cell growth (Horibata et al., 2015). shRNA targeting syntenin induced a 49% decrease in the growth of B16F10 cells, a 20 or 33% decrease in the growth of HT29 cells and a 44 or 46% decrease in the growth of MCF7 cells (respective values for the two different shRNA) (Figures 3C,D). Control shRNA, used as control, did not significantly affect colony formation, neither in plate nor in soft agar assays, when compared to empty vector transduced B16F10 and HT29 cells. Surprisingly, control shRNA significantly affected colony formation of MCF7 cells in both assays. The reason for this specific effect in MCF7 cells is unknown. While a priori this observation does not change the conclusion that syntenin depletion affects the growth of MCF7 cells, it indicates that from a quantitative point of view, data should be interpreted with caution.

Bottom Line: In human adulthood, syntenin gain-of-function is increasingly associated with various cancers and poor prognosis.We observed decreased migration, growth, and proliferation of the mouse melanoma cell line B16F10, the human colon cancer cell line HT29 and the human breast cancer cell line MCF7.We further documented that syntenin controls the presence of active β1 integrin at the cell membrane and G1/S cell cycle transition as well as the expression levels of CDK4, Cyclin D2, and Retinoblastoma proteins.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Signal Integration in Cell Fate Decision, Department of Human Genetics, KU Leuven Leuven, Belgium ; Centre de Recherche en Cancérologie de Marseille, Aix-Marseille Université Marseille, France ; Inserm U1068, Institut Paoli-Calmettes Marseille, France ; Centre National de la Recherche Scientifique, UMR7258 Marseille, France.

ABSTRACT
The scaffold protein syntenin abounds during fetal life where it is important for developmental movements. In human adulthood, syntenin gain-of-function is increasingly associated with various cancers and poor prognosis. Depending on the cancer model analyzed, syntenin affects various signaling pathways. We previously have shown that syntenin allows syndecan heparan sulfate proteoglycans to escape degradation. This indicates that syntenin has the potential to support sustained signaling of a plethora of growth factors and adhesion molecules. Here, we aim to clarify the impact of syntenin loss-of-function on cancer cell migration, growth, and proliferation, using cells from various cancer types and syntenin shRNA and siRNA silencing approaches. We observed decreased migration, growth, and proliferation of the mouse melanoma cell line B16F10, the human colon cancer cell line HT29 and the human breast cancer cell line MCF7. We further documented that syntenin controls the presence of active β1 integrin at the cell membrane and G1/S cell cycle transition as well as the expression levels of CDK4, Cyclin D2, and Retinoblastoma proteins. These data confirm that syntenin supports the migration and growth of tumor cells, independently of their origin, and further highlight the attractiveness of syntenin as potential therapeutic target.

No MeSH data available.


Related in: MedlinePlus