Phosphorylated c-Jun and Fra-1 induce matrix metalloproteinase-1 and thereby regulate invasion activity of 143B osteosarcoma cells

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Abstract

Osteosarcoma is the most common primary malignancy of bone and patients often develop pulmonary metastases. Despite the advances in surgical and medical management, the mechanisms underlying human osteosarcoma progression and metastasis remain to be elucidated. Gene expression profiles were compared by the cDNA microarray technique between two different human osteosarcoma sublines, MNNG/HOS and 143B, which differ greatly in spontaneous pulmonary metastatic potential. Here we report an enhanced expression of matrix metalloproteinase (MMP)-1 in the highly metastatic human osteosarcoma cell line 143B. Moreover, the in vitro invasion activity of 143B cells was MMP-1-dependent. The activator protein (AP)-1 binding site in the MMP-1 gene promoter was required for the constitutive expression of MMP-1 in 143B cells. Two AP-1 components, c-Jun and Fra-1, were phosphorylated, and bound to the AP-1 binding site of the MMP-1 promoter in 143B cells. Activated c-Jun and Fra-1 were essential for MMP-1 gene expression in 143B cells. Mitogen-activated protein kinase pathways including the c-Jun NH2-terminal kinase and the extracellular signal-regulated kinase activate c-Jun and Fra-1 and thereby regulate c-Jun/Fra-1 mediated events, establishing the mitogen-activated protein kinase/AP-1/MMP-1 axis as important in 143B cells. These data suggest that MMP-1 plays a central role in osteosarcoma invasion. Accordingly, MMP-1 might be a biomarker and therapeutic target for invasive osteosarcomas and pulmonary metastases.

Graphical abstract

Highlights

► Osteosarcoma patients develop pulmonary metastases, but its mechanisms remain unclear. ► MMP-1 and CXCR4 were notably expressed in a highly metastatic osteosarcoma cell line. ► MAPK pathways activated AP-1 (c-Jun/Fra-1), and thereby regulated MMP-1 expression. ► The MAPK/AP-1/MMP-1 axis plays a central role in osteosarcoma invasion.

Abbreviations

AP-1
activator protein-1
CXCR4
C–X–C chemokine receptor type 4
ECM
extracellular matrix
ELISA
enzyme-linked immunosorbent assay
EMSA
electrophoretic mobility shift assay
ERK
extracellular signal-regulated kinase
ETS
E-twenty six
FCS
fetal calf serum
Fra-1
Fos-related antigen-1
GAPDH
glyceraldehyde 3-phosphate dehydrogenase
IL
interleukin
IL-2Rα
IL-2 receptor α chain
JNK
c-Jun NH2-terminal kinase
MAPK
mitogen-activated protein kinase
MMP
matrix metalloproteinase
MNNG
N-methyl-N′-nitro-N-nitrosoguanidine
NF-κB
nuclear factor-κB
PBS
phosphate-buffered saline
PEA3
polyomavirus enhancer-A binding protein-3
Rb
retinoblastoma protein
RT-PCR
reverse transcription-PCR
SD
standard deviation
SDF-1α
stromal cell-derived factor-1α
siRNA
small interfering RNA
WST-8
water-soluble tetrazolium-8
WT
wild type

Keywords

MMP-1
Osteosarcoma
Invasion
Pulmonary metastasis
c-Jun
Fra-1

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