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The GPR 55 agonist, L-α-lysophosphatidylinositol, mediates ovarian carcinoma cell-induced angiogenesis.

Hofmann NA, Yang J, Trauger SA, Nakayama H, Huang L, Strunk D, Moses MA, Klagsbrun M, Bischoff J, Graier WF - Br. J. Pharmacol. (2015)

Bottom Line: To study underlying signal transduction, Western blot analysis was performed.Four target-specific siRNAs against GPR55 prevented these effects of LPI on angiogenesis.These pro-angiogenic effects of LPI were transduced by GPR55-dependent phosphorylation of ERK1/2 and p38 kinase.

View Article: PubMed Central - PubMed

Affiliation: Institute for Molecular Biology and Biochemistry, Medical University Graz, Graz, Austria.

No MeSH data available.


Related in: MedlinePlus

LPI stimulates angiogenesis in vitro and in vivo. (A–C) Effect of vehicle or 10 μM LPI on neonatal ECFC. (A) Cell numbers (×103) after 48 h in vitro proliferation assay. Dotted line marks starting cell number (12.000 cells). (B) Branch point formation in an in vitro angiogenesis assay after 16 h. (C) Closure of in vitro endothelial scratch area after 16 h. (A–C) Respective representative cell culture pictures with black bars marking 200 μm. n = 9; (D) Quantification of vessel numbers around white filter paper in an in vivo chicken CAM assay after 72 h with respective representative macroscopic pictures. n = 6–9; ***P < 0.001, significantly different from vehicle; Student's t-test.
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fig02: LPI stimulates angiogenesis in vitro and in vivo. (A–C) Effect of vehicle or 10 μM LPI on neonatal ECFC. (A) Cell numbers (×103) after 48 h in vitro proliferation assay. Dotted line marks starting cell number (12.000 cells). (B) Branch point formation in an in vitro angiogenesis assay after 16 h. (C) Closure of in vitro endothelial scratch area after 16 h. (A–C) Respective representative cell culture pictures with black bars marking 200 μm. n = 9; (D) Quantification of vessel numbers around white filter paper in an in vivo chicken CAM assay after 72 h with respective representative macroscopic pictures. n = 6–9; ***P < 0.001, significantly different from vehicle; Student's t-test.

Mentions: The effects of purified LPI on endothelial cell proliferation, network formation and migration were tested in vitro on isolated endothelial colony-forming progenitor cells (ECFCs) derived from three different donors. The isolated human neonatal cord ECFCs showed a distinct endothelial phenotype as shown by expression of typical endothelial cell surface markers (Supporting Information Fig. S1), as previously shown (Hofmann et al., 2009; 2012; Reinisch and Strunk, 2009; Reinisch et al., 2009). LPI stimulated ECFC proliferation in a dose-dependent manner with an EC50 of 2.8 (2.2–3.6) μM (Supporting Information Fig. S2a). Low concentrations of LPI, resembling the endogenous LPI levels secreted by 107 ovarian carcinoma cells (1 nM), were sufficient to stimulate proliferation of ECFCs (Supporting Information Fig. S2a). The maximum proliferative increase (1.55 ± 0.1-fold increase) was measured within 48 h upon applying 10 μM LPI as compared with vehicle controls (Figure 2A and Supporting Information Fig. S2a) and further experiments were performed at this 10 μM concentration. HUVECs showed a similar increase in proliferation as did isolated human adult peripheral blood ECFCs (Supporting Information Fig. S2b). Furthermore, compared with vehicle controls, 10 μM LPI significantly increased ECFC network formation in an in vitro Matrigel assay (Figure 2B) and closure of an endothelial wound in an in vitro scratch assay (Figure 2C).


The GPR 55 agonist, L-α-lysophosphatidylinositol, mediates ovarian carcinoma cell-induced angiogenesis.

Hofmann NA, Yang J, Trauger SA, Nakayama H, Huang L, Strunk D, Moses MA, Klagsbrun M, Bischoff J, Graier WF - Br. J. Pharmacol. (2015)

LPI stimulates angiogenesis in vitro and in vivo. (A–C) Effect of vehicle or 10 μM LPI on neonatal ECFC. (A) Cell numbers (×103) after 48 h in vitro proliferation assay. Dotted line marks starting cell number (12.000 cells). (B) Branch point formation in an in vitro angiogenesis assay after 16 h. (C) Closure of in vitro endothelial scratch area after 16 h. (A–C) Respective representative cell culture pictures with black bars marking 200 μm. n = 9; (D) Quantification of vessel numbers around white filter paper in an in vivo chicken CAM assay after 72 h with respective representative macroscopic pictures. n = 6–9; ***P < 0.001, significantly different from vehicle; Student's t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4543616&req=5

fig02: LPI stimulates angiogenesis in vitro and in vivo. (A–C) Effect of vehicle or 10 μM LPI on neonatal ECFC. (A) Cell numbers (×103) after 48 h in vitro proliferation assay. Dotted line marks starting cell number (12.000 cells). (B) Branch point formation in an in vitro angiogenesis assay after 16 h. (C) Closure of in vitro endothelial scratch area after 16 h. (A–C) Respective representative cell culture pictures with black bars marking 200 μm. n = 9; (D) Quantification of vessel numbers around white filter paper in an in vivo chicken CAM assay after 72 h with respective representative macroscopic pictures. n = 6–9; ***P < 0.001, significantly different from vehicle; Student's t-test.
Mentions: The effects of purified LPI on endothelial cell proliferation, network formation and migration were tested in vitro on isolated endothelial colony-forming progenitor cells (ECFCs) derived from three different donors. The isolated human neonatal cord ECFCs showed a distinct endothelial phenotype as shown by expression of typical endothelial cell surface markers (Supporting Information Fig. S1), as previously shown (Hofmann et al., 2009; 2012; Reinisch and Strunk, 2009; Reinisch et al., 2009). LPI stimulated ECFC proliferation in a dose-dependent manner with an EC50 of 2.8 (2.2–3.6) μM (Supporting Information Fig. S2a). Low concentrations of LPI, resembling the endogenous LPI levels secreted by 107 ovarian carcinoma cells (1 nM), were sufficient to stimulate proliferation of ECFCs (Supporting Information Fig. S2a). The maximum proliferative increase (1.55 ± 0.1-fold increase) was measured within 48 h upon applying 10 μM LPI as compared with vehicle controls (Figure 2A and Supporting Information Fig. S2a) and further experiments were performed at this 10 μM concentration. HUVECs showed a similar increase in proliferation as did isolated human adult peripheral blood ECFCs (Supporting Information Fig. S2b). Furthermore, compared with vehicle controls, 10 μM LPI significantly increased ECFC network formation in an in vitro Matrigel assay (Figure 2B) and closure of an endothelial wound in an in vitro scratch assay (Figure 2C).

Bottom Line: To study underlying signal transduction, Western blot analysis was performed.Four target-specific siRNAs against GPR55 prevented these effects of LPI on angiogenesis.These pro-angiogenic effects of LPI were transduced by GPR55-dependent phosphorylation of ERK1/2 and p38 kinase.

View Article: PubMed Central - PubMed

Affiliation: Institute for Molecular Biology and Biochemistry, Medical University Graz, Graz, Austria.

No MeSH data available.


Related in: MedlinePlus