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AP-1 as a Regulator of MMP-13 in the Stromal Cell of Giant Cell Tumor of Bone.

Mak IW, Turcotte RE, Popovic S, Singh G, Ghert M - Biochem Res Int (2011)

Bottom Line: We have previously shown that MMP-13 expression in these cells is regulated, at least in part, by the RUNX2 transcription factor.We then used siRNA gene knockdown to determine that these elements, in particular c-Jun, are upstream regulators of MMP-13 expression and activity in GCT stromal cells.We conclude that there was no synergy found between RUNX2 and AP-1 in the regulation of the MMP13 expression and that these transcription factors may be independently regulated in these cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, McMaster University, Hamilton, ON, Canada L8S 4L8.

ABSTRACT
Matrix-metalloproteinase-13 (MMP-13) has been shown to be an important protease in inflammatory and neoplastic conditions of the skeletal system. In particular, the stromal cells of giant cell tumor of bone (GCT) express very high levels of MMP-13 in response to the cytokine-rich environment of the tumor. We have previously shown that MMP-13 expression in these cells is regulated, at least in part, by the RUNX2 transcription factor. In the current study, we identify the expression of the c-Fos and c-Jun elements of the AP-1 transcription factor in these cells by protein screening assays and real-time PCR. We then used siRNA gene knockdown to determine that these elements, in particular c-Jun, are upstream regulators of MMP-13 expression and activity in GCT stromal cells. We conclude that there was no synergy found between RUNX2 and AP-1 in the regulation of the MMP13 expression and that these transcription factors may be independently regulated in these cells.

No MeSH data available.


Related in: MedlinePlus

Relative mRNA expression of the Runx2 and AP-1 transcription factors based on real-time RT-PCR. The expression of c-Jun, c-Fos, and Runx2 in GCT stromal cells treated with cytokines IL-1β for 24 h in serum-free media was analyzed using real-time PCR. The ΔΔCT method was used to calculate the real-time RT-PCR fold change using RPS18 mRNA for normalization, and all changes in expression are relative to the control without any treatment. Triplicate independent real-time PCR were performed.
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fig3: Relative mRNA expression of the Runx2 and AP-1 transcription factors based on real-time RT-PCR. The expression of c-Jun, c-Fos, and Runx2 in GCT stromal cells treated with cytokines IL-1β for 24 h in serum-free media was analyzed using real-time PCR. The ΔΔCT method was used to calculate the real-time RT-PCR fold change using RPS18 mRNA for normalization, and all changes in expression are relative to the control without any treatment. Triplicate independent real-time PCR were performed.

Mentions: To re-confirm the presence and expression of c-Fos and c-Jun of AP-1 in GCT stromal cells, the baseline mRNA expression level of c-Fos, c-Jun, and Runx2 was determined using real-time PCR. The mRNA expression of these three transcription factors was relatively low into the hundredth level relative to RPS18 expression (Figure 3). c-Fos had a similar expression level to that of Runx2, but c-Jun exhibited a 5-6-fold higher expression level.


AP-1 as a Regulator of MMP-13 in the Stromal Cell of Giant Cell Tumor of Bone.

Mak IW, Turcotte RE, Popovic S, Singh G, Ghert M - Biochem Res Int (2011)

Relative mRNA expression of the Runx2 and AP-1 transcription factors based on real-time RT-PCR. The expression of c-Jun, c-Fos, and Runx2 in GCT stromal cells treated with cytokines IL-1β for 24 h in serum-free media was analyzed using real-time PCR. The ΔΔCT method was used to calculate the real-time RT-PCR fold change using RPS18 mRNA for normalization, and all changes in expression are relative to the control without any treatment. Triplicate independent real-time PCR were performed.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3065034&req=5

fig3: Relative mRNA expression of the Runx2 and AP-1 transcription factors based on real-time RT-PCR. The expression of c-Jun, c-Fos, and Runx2 in GCT stromal cells treated with cytokines IL-1β for 24 h in serum-free media was analyzed using real-time PCR. The ΔΔCT method was used to calculate the real-time RT-PCR fold change using RPS18 mRNA for normalization, and all changes in expression are relative to the control without any treatment. Triplicate independent real-time PCR were performed.
Mentions: To re-confirm the presence and expression of c-Fos and c-Jun of AP-1 in GCT stromal cells, the baseline mRNA expression level of c-Fos, c-Jun, and Runx2 was determined using real-time PCR. The mRNA expression of these three transcription factors was relatively low into the hundredth level relative to RPS18 expression (Figure 3). c-Fos had a similar expression level to that of Runx2, but c-Jun exhibited a 5-6-fold higher expression level.

Bottom Line: We have previously shown that MMP-13 expression in these cells is regulated, at least in part, by the RUNX2 transcription factor.We then used siRNA gene knockdown to determine that these elements, in particular c-Jun, are upstream regulators of MMP-13 expression and activity in GCT stromal cells.We conclude that there was no synergy found between RUNX2 and AP-1 in the regulation of the MMP13 expression and that these transcription factors may be independently regulated in these cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, McMaster University, Hamilton, ON, Canada L8S 4L8.

ABSTRACT
Matrix-metalloproteinase-13 (MMP-13) has been shown to be an important protease in inflammatory and neoplastic conditions of the skeletal system. In particular, the stromal cells of giant cell tumor of bone (GCT) express very high levels of MMP-13 in response to the cytokine-rich environment of the tumor. We have previously shown that MMP-13 expression in these cells is regulated, at least in part, by the RUNX2 transcription factor. In the current study, we identify the expression of the c-Fos and c-Jun elements of the AP-1 transcription factor in these cells by protein screening assays and real-time PCR. We then used siRNA gene knockdown to determine that these elements, in particular c-Jun, are upstream regulators of MMP-13 expression and activity in GCT stromal cells. We conclude that there was no synergy found between RUNX2 and AP-1 in the regulation of the MMP13 expression and that these transcription factors may be independently regulated in these cells.

No MeSH data available.


Related in: MedlinePlus