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Functional consequences of prolactin signalling in endothelial cells: a potential link with angiogenesis in pathophysiology?

Reuwer AQ, Nowak-Sliwinska P, Mans LA, van der Loos CM, von der Thüsen JH, Twickler MT, Spek CA, Goffin V, Griffioen AW, Borensztajn KS - J. Cell. Mol. Med. (2012)

Bottom Line: These effects are blocked by a specific prolactin receptor antagonist, del1-9-G129R-hPRL.Interestingly, while prolactin has only little effect on endothelial cell proliferation, it markedly stimulates endothelial cell migration.Again, migration was reverted by del1-9-G129R-hPRL, indicating a direct effect of prolactin on its receptor.

View Article: PubMed Central - PubMed

Affiliation: Department of Vascular Medicine, Academic Medical Center, Amsterdam, The Netherlands. a.q.reuwer@amc.uva.nl

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Related in: MedlinePlus

Prolactin exerts a direct effect on endothelial cells. 2H11 endothelial cells, serum starved for 8 hrs, were stimulated for 30 min. with prolactin (10, 100, 500 or 1000 μg/l; lane 5–9) or with prolactin (100 or 500 μg/l) in combination with the prolactin receptor antagonist del1-9-G129R-hPRL (‘Anta’; 2000 or 10,000 μg/l; lane 1 and 2) or with del1-9-G129R-hPRL alone (2000 or 10,000 μg/l; lane 3 and 4). Cell lysates were analysed using Western blot for phosphorylated STAT5 (‘Phospho-STAT5’; top panel), phosphorylated ERK1/2 (‘Phospho-ERK1/2’; middle panel) and tubulin as loading control (bottom panel). A representative picture of an experiment is shown, which was performed three times with similar results.
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fig01: Prolactin exerts a direct effect on endothelial cells. 2H11 endothelial cells, serum starved for 8 hrs, were stimulated for 30 min. with prolactin (10, 100, 500 or 1000 μg/l; lane 5–9) or with prolactin (100 or 500 μg/l) in combination with the prolactin receptor antagonist del1-9-G129R-hPRL (‘Anta’; 2000 or 10,000 μg/l; lane 1 and 2) or with del1-9-G129R-hPRL alone (2000 or 10,000 μg/l; lane 3 and 4). Cell lysates were analysed using Western blot for phosphorylated STAT5 (‘Phospho-STAT5’; top panel), phosphorylated ERK1/2 (‘Phospho-ERK1/2’; middle panel) and tubulin as loading control (bottom panel). A representative picture of an experiment is shown, which was performed three times with similar results.

Mentions: Upon activation of the prolactin receptor, the JAK-STAT signalling pathway is activated, with STAT5 as the primary mediator of prolactin action [2]. Additionally, ERK1/2 phosphorylation is also a major downstream target of prolactin receptor signalling [2]. Hence, we investigated the phosphorylation of STAT5 and ERK1/2 after exposure of 2H11 cells to a dose range of prolactin. As shown in Figure 1, prolactin induced a concentration-dependent phosphorylation of ERK1/2, starting at 10 μg/l and culminating at 1000 μg/l (lane 5–8). Phosphorylation of ERK1/2 was decreased when using 10,000 μg/l prolactin (lane 9). Prolactin also induced the phosphorylation of STAT5, starting at 10 μg/l and peaking at 10,000 μg/l (lane 5–9). Moreover, ERK1/2 and STAT5 phosphorylation induced by prolactin was completely abolished by addition of 20-fold molar excess of the prolactin receptor antagonist del1-9-G129R-hPRL (lane 1 and 2 compared to 6 and 7). The antagonist alone, even at extremely high concentrations (10,000 μg/l), failed to notably induce signalling (lane 3 and 4) (Fig. 1). Overall, these data demonstrate that prolactin has a direct effect on endothelial cells, which is mediated by the prolactin receptor.


Functional consequences of prolactin signalling in endothelial cells: a potential link with angiogenesis in pathophysiology?

Reuwer AQ, Nowak-Sliwinska P, Mans LA, van der Loos CM, von der Thüsen JH, Twickler MT, Spek CA, Goffin V, Griffioen AW, Borensztajn KS - J. Cell. Mol. Med. (2012)

Prolactin exerts a direct effect on endothelial cells. 2H11 endothelial cells, serum starved for 8 hrs, were stimulated for 30 min. with prolactin (10, 100, 500 or 1000 μg/l; lane 5–9) or with prolactin (100 or 500 μg/l) in combination with the prolactin receptor antagonist del1-9-G129R-hPRL (‘Anta’; 2000 or 10,000 μg/l; lane 1 and 2) or with del1-9-G129R-hPRL alone (2000 or 10,000 μg/l; lane 3 and 4). Cell lysates were analysed using Western blot for phosphorylated STAT5 (‘Phospho-STAT5’; top panel), phosphorylated ERK1/2 (‘Phospho-ERK1/2’; middle panel) and tubulin as loading control (bottom panel). A representative picture of an experiment is shown, which was performed three times with similar results.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3822974&req=5

fig01: Prolactin exerts a direct effect on endothelial cells. 2H11 endothelial cells, serum starved for 8 hrs, were stimulated for 30 min. with prolactin (10, 100, 500 or 1000 μg/l; lane 5–9) or with prolactin (100 or 500 μg/l) in combination with the prolactin receptor antagonist del1-9-G129R-hPRL (‘Anta’; 2000 or 10,000 μg/l; lane 1 and 2) or with del1-9-G129R-hPRL alone (2000 or 10,000 μg/l; lane 3 and 4). Cell lysates were analysed using Western blot for phosphorylated STAT5 (‘Phospho-STAT5’; top panel), phosphorylated ERK1/2 (‘Phospho-ERK1/2’; middle panel) and tubulin as loading control (bottom panel). A representative picture of an experiment is shown, which was performed three times with similar results.
Mentions: Upon activation of the prolactin receptor, the JAK-STAT signalling pathway is activated, with STAT5 as the primary mediator of prolactin action [2]. Additionally, ERK1/2 phosphorylation is also a major downstream target of prolactin receptor signalling [2]. Hence, we investigated the phosphorylation of STAT5 and ERK1/2 after exposure of 2H11 cells to a dose range of prolactin. As shown in Figure 1, prolactin induced a concentration-dependent phosphorylation of ERK1/2, starting at 10 μg/l and culminating at 1000 μg/l (lane 5–8). Phosphorylation of ERK1/2 was decreased when using 10,000 μg/l prolactin (lane 9). Prolactin also induced the phosphorylation of STAT5, starting at 10 μg/l and peaking at 10,000 μg/l (lane 5–9). Moreover, ERK1/2 and STAT5 phosphorylation induced by prolactin was completely abolished by addition of 20-fold molar excess of the prolactin receptor antagonist del1-9-G129R-hPRL (lane 1 and 2 compared to 6 and 7). The antagonist alone, even at extremely high concentrations (10,000 μg/l), failed to notably induce signalling (lane 3 and 4) (Fig. 1). Overall, these data demonstrate that prolactin has a direct effect on endothelial cells, which is mediated by the prolactin receptor.

Bottom Line: These effects are blocked by a specific prolactin receptor antagonist, del1-9-G129R-hPRL.Interestingly, while prolactin has only little effect on endothelial cell proliferation, it markedly stimulates endothelial cell migration.Again, migration was reverted by del1-9-G129R-hPRL, indicating a direct effect of prolactin on its receptor.

View Article: PubMed Central - PubMed

Affiliation: Department of Vascular Medicine, Academic Medical Center, Amsterdam, The Netherlands. a.q.reuwer@amc.uva.nl

Show MeSH
Related in: MedlinePlus