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Effect of fibroblast growth factor 9 on Runx2 gene promoter activity in MC3T3-E1 and C2C12 cells
Li-yun Yu1, Yu Pei, Wei-bo Xia
1Department of Anesthesiology, Beijing Electric Power Hospital, Beijing 100073, China.
Background:
Fibroblast growth factor 9 (FGF9), expressed in brain, kidney and developing skeletal tissues, can physiologically inhibit endochondral ossification; but little is known about how FGF9 affects osteoblasts and its detailed regulatory mechanism. Here we examined the effect of FGF9 on the activity of the murine Runt-related transcription factor 2 (Runx2) gene promoter in preosteoblast MC3T3-E1 and premyoblast C2C12 cells.
Methods:
Plasmids containing the Runx2 promoter region were transfected into MC3T3-E1 and C2C12 cells and stably transfected cell lines were established. The method of luciferase reporter gene activation was used to examine the effects of FGF9 on the promoter activity.
Results:
FGF9 (10 ng/ml) increased Runx2 promoter activity in MC3T3-E1 cells. When MC3T3-E1 cells were treated with FGF9 plus the various inhibitors or activator of the intracellular signaling transducation pathways, including 10 micromol/L U0126 (the inhibitor of mitogen-activated protein kinase kinase), 10 micromol/L SB203580 (the inhibitor of p38/mitogen activated protein kinase), or 1 micromol/L C6 ceramide (an activator of mitogen activated protein kinase), the luciferase expression did not change significantly compared with that of the cells treated with FGF9 only. However, when C2C12 cells were treated with 10 ng/ml FGF9, Runx2 gene promoter activity first decreased and then increased over a period of 1 to 5 days. Among the above inhibitors, only U0126 (10 micromol/L) completely blocked the effects of FGF9 on Runx2 gene promoter activity.
Conclusions:
Our data showed that FGF9 can affect Runx2 gene promoter activity in MC3T3-E1 and C2C12 cells. The action of FGF9 appears to depend partly on the mitogen-activated protein kinase kinase/mitogen-activated protein kinase pathways in C2C12 cells.
Insights
Fibroblast growth factor 9 (FGF9) influences Runt-related transcription factor 2 (Runx2) gene promoter activity in bone and muscle cells. FGF9
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Fibroblast growth factor 9 (FGF9) plays a role in skeletal development and can inhibit ossification.
- The precise mechanisms by which FGF9 impacts osteoblasts remain largely unexplored.
- This study investigates FGF9's effect on the Runt-related transcription factor 2 (Runx2) gene promoter.
Purpose of the Study:
- To elucidate the regulatory role of FGF9 in osteoblast activity.
- To examine the impact of FGF9 on the murine Runx2 gene promoter.
- To identify signaling pathways involved in FGF9's action on Runx2.
Main Methods:
- Stable transfection of MC3T3-E1 (preosteoblast) and C2C12 (premyoblast) cell lines with Runx2 promoter plasmids.
- Utilized luciferase reporter gene assays to quantify promoter activity.
- Administered FGF9 and various pathway inhibitors/activators (U0126, SB203580, C6 ceramide) to assess signaling mechanisms.
Main Results:
- FGF9 (10 ng/ml) enhanced Runx2 promoter activity in MC3T3-E1 cells.
- In C2C12 cells, FGF9 initially decreased then increased Runx2 promoter activity over 1-5 days.
- The mitogen-activated protein kinase kinase/mitogen-activated protein kinase pathway inhibitor U0126 blocked FGF9's effect on Runx2 in C2C12 cells.
Conclusions:
- FGF9 modulates Runx2 gene promoter activity in both preosteoblast and premyoblast cell lines.
- The observed effects of FGF9 on Runx2 promoter activity in C2C12 cells are partially mediated by the MAPK signaling pathway.
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