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Survivin is an essential mediator of arthritis interacting with urokinase signalling.

Baran M, Möllers LN, Andersson S, Jonsson IM, Ekwall AK, Bjersing J, Tarkowski A, Bokarewa M - J. Cell. Mol. Med. (2009)

Bottom Line: Using inhibitors of intracellular signalling, we show that uPA and survivin share common transduction pathways in synovial fibroblasts being dependent on the activity of tyrosine kinases, phosphatidylinositide 3 kinase and mitogen effector kinase.Moreover, uPA production is significantly reduced in fibroblasts if survivin synthesis has been silenced by siRNA.Close correlation between survivin and uPA levels in patients with RA supports the importance of this connection for the pathogenesis of arthritis.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology and Inflammation Research, Sahlgrenska University Hospital, Gothenburg, Sweden.

ABSTRACT
Proto-oncogene survivin has recently been identified as a prognostic marker distinguishing patients with destructive rheumatoid arthritis (RA). In the present material of 132 RA patients and 82 controls, the levels of survivin correlated to urokinase (uPA) (r= 0.46), a plasminogen activator over-expressed in inflamed joints and known to exhibit potent arthritogenic properties. Here we evaluate the functional relationship between these proteins using primary synovial fibroblasts and leucocytes of RA patients, human monocytic (THP-1) and fibroblast (MRC-5) cell lines. Using inhibitors of intracellular signalling, we show that uPA and survivin share common transduction pathways in synovial fibroblasts being dependent on the activity of tyrosine kinases, phosphatidylinositide 3 kinase and mitogen effector kinase. Moreover, uPA production is significantly reduced in fibroblasts if survivin synthesis has been silenced by siRNA. Importantly, silencing of survivin in fibroblasts prevented their invasive growth in knee joints of severe combined immune deficient mice. Interaction of uPA with receptor up-regulates survivin expression in leucocytes. In turn, survivin is required for the up-regulation of uPA receptor on the cell surface. These findings indicate that survivin is an essential mediator of arthritogenic properties of uPA regulating its synthesis in synovial fibroblasts and uPAR expression in leucocytes. Close correlation between survivin and uPA levels in patients with RA supports the importance of this connection for the pathogenesis of arthritis.

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uPA up-regulates production of survivin in human leucocytes. Cells were seeded in concentration of 1 × 106/ml in 96-well plates and incubated for 48 hrs with uPA at final concentrations of 0-10-100-1000 UI/ml (PBMC, n= 5, A), and 0-30-300 UI/ml (THP-1, n= 13, B). Following 48 hrs, survivin levels were evaluated in the cell lysates by an ELISA. Results are presented as the increase in the ratio of survivin levels in uPA-stimulated cells in comparison to non-stimulated cell cultures. In addition, the proliferation rates in uPA-stimulated PBMC cultures are presented (A, open columns).
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fig04: uPA up-regulates production of survivin in human leucocytes. Cells were seeded in concentration of 1 × 106/ml in 96-well plates and incubated for 48 hrs with uPA at final concentrations of 0-10-100-1000 UI/ml (PBMC, n= 5, A), and 0-30-300 UI/ml (THP-1, n= 13, B). Following 48 hrs, survivin levels were evaluated in the cell lysates by an ELISA. Results are presented as the increase in the ratio of survivin levels in uPA-stimulated cells in comparison to non-stimulated cell cultures. In addition, the proliferation rates in uPA-stimulated PBMC cultures are presented (A, open columns).

Mentions: To investigate the functional relationship between uPA and survivin in human leucocytes, PBMCs from healthy individuals (n= 5) were stimulated with uPA (0, 10, 100, 1000 UI/ml). The levels of survivin were evaluated in cell lysates 48 hrs following stimulation. A dose-dependent intracellular up-regulation of survivin production was observed (Fig. 4A). A relation of survivin expression to the proliferative capacity of uPA was measured by [6-3H]-thymidine intake of the uPA-stimulated PBMCs (Fig. 4A). This induction of survivin level in PBMCs was only partly related to proliferation of cells. The uPA stimulation had a similar effect on THP-1 cells, a human monocytic cell line. In concordance with the effect on PBMCs, THP-1 cells dose-dependently increased survivin production in response to uPA as compared with non-stimulated cells (in fold increase, 1.80 ± 0.15, P= 0.0002) (Fig. 4B).


Survivin is an essential mediator of arthritis interacting with urokinase signalling.

Baran M, Möllers LN, Andersson S, Jonsson IM, Ekwall AK, Bjersing J, Tarkowski A, Bokarewa M - J. Cell. Mol. Med. (2009)

uPA up-regulates production of survivin in human leucocytes. Cells were seeded in concentration of 1 × 106/ml in 96-well plates and incubated for 48 hrs with uPA at final concentrations of 0-10-100-1000 UI/ml (PBMC, n= 5, A), and 0-30-300 UI/ml (THP-1, n= 13, B). Following 48 hrs, survivin levels were evaluated in the cell lysates by an ELISA. Results are presented as the increase in the ratio of survivin levels in uPA-stimulated cells in comparison to non-stimulated cell cultures. In addition, the proliferation rates in uPA-stimulated PBMC cultures are presented (A, open columns).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4516528&req=5

fig04: uPA up-regulates production of survivin in human leucocytes. Cells were seeded in concentration of 1 × 106/ml in 96-well plates and incubated for 48 hrs with uPA at final concentrations of 0-10-100-1000 UI/ml (PBMC, n= 5, A), and 0-30-300 UI/ml (THP-1, n= 13, B). Following 48 hrs, survivin levels were evaluated in the cell lysates by an ELISA. Results are presented as the increase in the ratio of survivin levels in uPA-stimulated cells in comparison to non-stimulated cell cultures. In addition, the proliferation rates in uPA-stimulated PBMC cultures are presented (A, open columns).
Mentions: To investigate the functional relationship between uPA and survivin in human leucocytes, PBMCs from healthy individuals (n= 5) were stimulated with uPA (0, 10, 100, 1000 UI/ml). The levels of survivin were evaluated in cell lysates 48 hrs following stimulation. A dose-dependent intracellular up-regulation of survivin production was observed (Fig. 4A). A relation of survivin expression to the proliferative capacity of uPA was measured by [6-3H]-thymidine intake of the uPA-stimulated PBMCs (Fig. 4A). This induction of survivin level in PBMCs was only partly related to proliferation of cells. The uPA stimulation had a similar effect on THP-1 cells, a human monocytic cell line. In concordance with the effect on PBMCs, THP-1 cells dose-dependently increased survivin production in response to uPA as compared with non-stimulated cells (in fold increase, 1.80 ± 0.15, P= 0.0002) (Fig. 4B).

Bottom Line: Using inhibitors of intracellular signalling, we show that uPA and survivin share common transduction pathways in synovial fibroblasts being dependent on the activity of tyrosine kinases, phosphatidylinositide 3 kinase and mitogen effector kinase.Moreover, uPA production is significantly reduced in fibroblasts if survivin synthesis has been silenced by siRNA.Close correlation between survivin and uPA levels in patients with RA supports the importance of this connection for the pathogenesis of arthritis.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology and Inflammation Research, Sahlgrenska University Hospital, Gothenburg, Sweden.

ABSTRACT
Proto-oncogene survivin has recently been identified as a prognostic marker distinguishing patients with destructive rheumatoid arthritis (RA). In the present material of 132 RA patients and 82 controls, the levels of survivin correlated to urokinase (uPA) (r= 0.46), a plasminogen activator over-expressed in inflamed joints and known to exhibit potent arthritogenic properties. Here we evaluate the functional relationship between these proteins using primary synovial fibroblasts and leucocytes of RA patients, human monocytic (THP-1) and fibroblast (MRC-5) cell lines. Using inhibitors of intracellular signalling, we show that uPA and survivin share common transduction pathways in synovial fibroblasts being dependent on the activity of tyrosine kinases, phosphatidylinositide 3 kinase and mitogen effector kinase. Moreover, uPA production is significantly reduced in fibroblasts if survivin synthesis has been silenced by siRNA. Importantly, silencing of survivin in fibroblasts prevented their invasive growth in knee joints of severe combined immune deficient mice. Interaction of uPA with receptor up-regulates survivin expression in leucocytes. In turn, survivin is required for the up-regulation of uPA receptor on the cell surface. These findings indicate that survivin is an essential mediator of arthritogenic properties of uPA regulating its synthesis in synovial fibroblasts and uPAR expression in leucocytes. Close correlation between survivin and uPA levels in patients with RA supports the importance of this connection for the pathogenesis of arthritis.

Show MeSH
Related in: MedlinePlus