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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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Related in: MedlinePlus

Effects of C/EBPβ and S100A4 expression on cell proliferationThe proliferation of mouse (C1, I4 and I5) and human (TC and TI) cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector was determined using “QIA127 Rapid Cell Proliferation Kit”, as indicated in Methods. Cells were seeded into individual wells of a 96-well plate and cultivated for 24 h after which WST-1 was added to the culture medium. The cleavage of the WST-1 was quantified by absorbance measurement at 450 nm. Values are the means ± S.D. of three different experiments in triplicate.
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Figure 7: Effects of C/EBPβ and S100A4 expression on cell proliferationThe proliferation of mouse (C1, I4 and I5) and human (TC and TI) cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector was determined using “QIA127 Rapid Cell Proliferation Kit”, as indicated in Methods. Cells were seeded into individual wells of a 96-well plate and cultivated for 24 h after which WST-1 was added to the culture medium. The cleavage of the WST-1 was quantified by absorbance measurement at 450 nm. Values are the means ± S.D. of three different experiments in triplicate.

Mentions: In order to know whether S100A4 could also play a role in proliferation of GL261 and T98G cells, we performed “QIA127 Rapid Cell Proliferation Kit” assay both in control and C/EBPβ-depleted cells when S100A4 is overexpressed. As can be shown in Figure 7, we didn't find any difference in growth and viability between control and S100A4-overexpressing cells. These results are in agreement with data from Takenaga et al [35] showing that S100A4 is not involved in the growth of C6 glioblastoma 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)

Effects of C/EBPβ and S100A4 expression on cell proliferationThe proliferation of mouse (C1, I4 and I5) and human (TC and TI) cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector was determined using “QIA127 Rapid Cell Proliferation Kit”, as indicated in Methods. Cells were seeded into individual wells of a 96-well plate and cultivated for 24 h after which WST-1 was added to the culture medium. The cleavage of the WST-1 was quantified by absorbance measurement at 450 nm. Values are the means ± S.D. of three different experiments in triplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 7: Effects of C/EBPβ and S100A4 expression on cell proliferationThe proliferation of mouse (C1, I4 and I5) and human (TC and TI) cells transfected with the S100A4 expressing vector pIRES2-DsRed-Express or the corresponding control vector was determined using “QIA127 Rapid Cell Proliferation Kit”, as indicated in Methods. Cells were seeded into individual wells of a 96-well plate and cultivated for 24 h after which WST-1 was added to the culture medium. The cleavage of the WST-1 was quantified by absorbance measurement at 450 nm. Values are the means ± S.D. of three different experiments in triplicate.
Mentions: In order to know whether S100A4 could also play a role in proliferation of GL261 and T98G cells, we performed “QIA127 Rapid Cell Proliferation Kit” assay both in control and C/EBPβ-depleted cells when S100A4 is overexpressed. As can be shown in Figure 7, we didn't find any difference in growth and viability between control and S100A4-overexpressing cells. These results are in agreement with data from Takenaga et al [35] showing that S100A4 is not involved in the growth of C6 glioblastoma 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