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CCAAT/enhancer binding protein β induces motility and invasion of glioblastoma cells through transcriptional regulation of the calcium binding protein S100A4.

Aguilar-Morante D, Morales-Garcia JA, Santos A, Perez-Castillo A - Oncotarget (2015)

Bottom Line: In agreement with this, we showed that C/EBPβ depletion decreases the mRNA levels of different genes involved in metastasis and invasion.Our results show that C/EBPβ suppression significantly reduced the levels of S100A4 in murine GL261 and human T98G glioblastoma cells.By employing an S100A4-promoter reporter, we observed a significant induction in the transcriptional activation of the S100A4 gene by C/EBPβ.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigaciones Biomédicas, (CSIC-UAM), Departamento Modelos Experimentales de Enfermedades Humanas, Arturo Duperier, Madrid, Spain.

ABSTRACT
We have previously shown that decreased expression of CCAAT/enhancer binding protein β (C/EBPβ) inhibits the growth of glioblastoma cells and diminishes their transformation capacity and migration. In agreement with this, we showed that C/EBPβ depletion decreases the mRNA levels of different genes involved in metastasis and invasion. Among these, we found S100 calcium binding protein A4 (S100A4) to be almost undetectable in glioblastoma cells deficient in C/EBPβ. Here, we have evaluated the possible role of S100A4 in the observed effects of C/EBPβ in glioblastoma cells and the mechanism through which S100A4 levels are controlled by C/EBPβ. Our results show that C/EBPβ suppression significantly reduced the levels of S100A4 in murine GL261 and human T98G glioblastoma cells. By employing an S100A4-promoter reporter, we observed a significant induction in the transcriptional activation of the S100A4 gene by C/EBPβ. Furthermore, overexpression of S100A4 in C/EBPβ-depleted glioblastoma cells reverses the enhanced migration and motility induced by this transcription factor. Our data also point to a role of S100A4 in glioblastoma cell invasion and suggest that the C/EBPβ gene controls the invasive potential of GL261 and T98G cells through direct regulation of S100A4. Finally, this study indicates a role of C/EBPβ on the maintenance of the stem cell population present in GL261 glioblastoma cells.

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Effect of C/EBPβ and S100A4 on T98G cells motilityTC and TI cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector were grown until reach confluence. A linear scratch was performed with a plastic pipette tip. Images were taken with a phase contrast microscope at different times after wounding. (A) Representative images and (B) quantifications of the in vitro wound-healing assay are shown. Bar scale 100 μm. ***p < 0.001; **p < 0.01.
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Figure 6: Effect of C/EBPβ and S100A4 on T98G cells motilityTC and TI cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector were grown until reach confluence. A linear scratch was performed with a plastic pipette tip. Images were taken with a phase contrast microscope at different times after wounding. (A) Representative images and (B) quantifications of the in vitro wound-healing assay are shown. Bar scale 100 μm. ***p < 0.001; **p < 0.01.

Mentions: Next, we determined the effect of C/EBPβ depletion and S100A4 overexpression (Figure 5A and 5B) on invasion and motility in the human glioblastoma cell line T98G. Similar to the result observed in GL261 cells, C/EBPβ depletion in T98G cells caused a marked decrease in S100A4 protein content (Figure 5A), invasion capacity (Figure 5C and 5D) and cell motility (Figure 6). The overexpression of S100A4 in T98G cells, as in GL261 cells, increased invasion capacity (Figure 5C) and motility (Figure 6A and 6B) in both non C/EBPβ-depleted (TC) and C/EBPβ-depleted (TI) cells.


CCAAT/enhancer binding protein β induces motility and invasion of glioblastoma cells through transcriptional regulation of the calcium binding protein S100A4.

Aguilar-Morante D, Morales-Garcia JA, Santos A, Perez-Castillo A - Oncotarget (2015)

Effect of C/EBPβ and S100A4 on T98G cells motilityTC and TI cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector were grown until reach confluence. A linear scratch was performed with a plastic pipette tip. Images were taken with a phase contrast microscope at different times after wounding. (A) Representative images and (B) quantifications of the in vitro wound-healing assay are shown. Bar scale 100 μm. ***p < 0.001; **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414196&req=5

Figure 6: Effect of C/EBPβ and S100A4 on T98G cells motilityTC and TI cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector were grown until reach confluence. A linear scratch was performed with a plastic pipette tip. Images were taken with a phase contrast microscope at different times after wounding. (A) Representative images and (B) quantifications of the in vitro wound-healing assay are shown. Bar scale 100 μm. ***p < 0.001; **p < 0.01.
Mentions: Next, we determined the effect of C/EBPβ depletion and S100A4 overexpression (Figure 5A and 5B) on invasion and motility in the human glioblastoma cell line T98G. Similar to the result observed in GL261 cells, C/EBPβ depletion in T98G cells caused a marked decrease in S100A4 protein content (Figure 5A), invasion capacity (Figure 5C and 5D) and cell motility (Figure 6). The overexpression of S100A4 in T98G cells, as in GL261 cells, increased invasion capacity (Figure 5C) and motility (Figure 6A and 6B) in both non C/EBPβ-depleted (TC) and C/EBPβ-depleted (TI) cells.

Bottom Line: In agreement with this, we showed that C/EBPβ depletion decreases the mRNA levels of different genes involved in metastasis and invasion.Our results show that C/EBPβ suppression significantly reduced the levels of S100A4 in murine GL261 and human T98G glioblastoma cells.By employing an S100A4-promoter reporter, we observed a significant induction in the transcriptional activation of the S100A4 gene by C/EBPβ.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigaciones Biomédicas, (CSIC-UAM), Departamento Modelos Experimentales de Enfermedades Humanas, Arturo Duperier, Madrid, Spain.

ABSTRACT
We have previously shown that decreased expression of CCAAT/enhancer binding protein β (C/EBPβ) inhibits the growth of glioblastoma cells and diminishes their transformation capacity and migration. In agreement with this, we showed that C/EBPβ depletion decreases the mRNA levels of different genes involved in metastasis and invasion. Among these, we found S100 calcium binding protein A4 (S100A4) to be almost undetectable in glioblastoma cells deficient in C/EBPβ. Here, we have evaluated the possible role of S100A4 in the observed effects of C/EBPβ in glioblastoma cells and the mechanism through which S100A4 levels are controlled by C/EBPβ. Our results show that C/EBPβ suppression significantly reduced the levels of S100A4 in murine GL261 and human T98G glioblastoma cells. By employing an S100A4-promoter reporter, we observed a significant induction in the transcriptional activation of the S100A4 gene by C/EBPβ. Furthermore, overexpression of S100A4 in C/EBPβ-depleted glioblastoma cells reverses the enhanced migration and motility induced by this transcription factor. Our data also point to a role of S100A4 in glioblastoma cell invasion and suggest that the C/EBPβ gene controls the invasive potential of GL261 and T98G cells through direct regulation of S100A4. Finally, this study indicates a role of C/EBPβ on the maintenance of the stem cell population present in GL261 glioblastoma cells.

Show MeSH
Related in: MedlinePlus