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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

Ovarian cancer cells produce LPI and induce chicken CAM angiogenesis in a GPR55-dependent manner. (A) Quantification of LPI in conditioned medium from three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362). (B) Quantification of vessel numbers around white filter paper in an in vivo CAM assay (by ImageJ). Filter papers were loaded with unconditioned DMEM or 24 h conditioned DMEM (CM) of three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362), respectively, with or without vehicle or GPR55 inhibitor CID16020046 (CID). Representative macroscopic pictures of CAM angiogenesis around filter paper containing control DMEM, OVCAR-5 CM or OVCAR-5 CM with CID. n = 6–9; *P < 0.05; **P < 0.01, significantly different from vehicle control; #P < 0.01, significantly different from corresponding ovarian cancer CM. anova followed by Bonferroni test.
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fig01: Ovarian cancer cells produce LPI and induce chicken CAM angiogenesis in a GPR55-dependent manner. (A) Quantification of LPI in conditioned medium from three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362). (B) Quantification of vessel numbers around white filter paper in an in vivo CAM assay (by ImageJ). Filter papers were loaded with unconditioned DMEM or 24 h conditioned DMEM (CM) of three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362), respectively, with or without vehicle or GPR55 inhibitor CID16020046 (CID). Representative macroscopic pictures of CAM angiogenesis around filter paper containing control DMEM, OVCAR-5 CM or OVCAR-5 CM with CID. n = 6–9; *P < 0.05; **P < 0.01, significantly different from vehicle control; #P < 0.01, significantly different from corresponding ovarian cancer CM. anova followed by Bonferroni test.

Mentions: Increased serum levels of the GPR55-ligand LPI have been found in patients with high-grade ovarian carcinoma (Xiao et al., 2000; 2001; Xu et al., 2001; Sutphen et al., 2004; Murph et al., 2007; Pineiro et al., 2011; Pineiro and Falasca, 2012). To test our hypothesis, that ovarian cancer cells secrete LPI, and thus promote tumour angiogenesis in vivo via an LPI/GPR55-dependent mechanism; conditioned medium from the human ovarian cancer cell lines OVCAR-3, OVCAR-5 and COV-362 was analysed for its LPI levels and in the CAM angiogenesis model. LC-MS/MS revealed that OVCAR-3, OVCAR-5 and COV-362 cells produced significant but quite different amounts of LPI (Figure 1A). Within 3 days, conditioned medium from OVCAR-3, OVCAR-5 and COV-362 strongly induced angiogenesis in vivo to a similar extent (90–100% increase), compared with unconditioned medium (Figure 1B). Selective inhibition of the LPI receptor GPR55 with CID16020046 (20 μM) effectively blocked ovarian cancer-induced angiogenesis of all tested cell lines (Figure 1B). Together, these results suggest that LPI produced by ovarian cancer cells induces angiogenesis in a GPR55-dependent manner.


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)

Ovarian cancer cells produce LPI and induce chicken CAM angiogenesis in a GPR55-dependent manner. (A) Quantification of LPI in conditioned medium from three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362). (B) Quantification of vessel numbers around white filter paper in an in vivo CAM assay (by ImageJ). Filter papers were loaded with unconditioned DMEM or 24 h conditioned DMEM (CM) of three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362), respectively, with or without vehicle or GPR55 inhibitor CID16020046 (CID). Representative macroscopic pictures of CAM angiogenesis around filter paper containing control DMEM, OVCAR-5 CM or OVCAR-5 CM with CID. n = 6–9; *P < 0.05; **P < 0.01, significantly different from vehicle control; #P < 0.01, significantly different from corresponding ovarian cancer CM. anova followed by Bonferroni test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig01: Ovarian cancer cells produce LPI and induce chicken CAM angiogenesis in a GPR55-dependent manner. (A) Quantification of LPI in conditioned medium from three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362). (B) Quantification of vessel numbers around white filter paper in an in vivo CAM assay (by ImageJ). Filter papers were loaded with unconditioned DMEM or 24 h conditioned DMEM (CM) of three different ovarian cancer cell lines (OVCAR-3, OVCAR-5, COV-362), respectively, with or without vehicle or GPR55 inhibitor CID16020046 (CID). Representative macroscopic pictures of CAM angiogenesis around filter paper containing control DMEM, OVCAR-5 CM or OVCAR-5 CM with CID. n = 6–9; *P < 0.05; **P < 0.01, significantly different from vehicle control; #P < 0.01, significantly different from corresponding ovarian cancer CM. anova followed by Bonferroni test.
Mentions: Increased serum levels of the GPR55-ligand LPI have been found in patients with high-grade ovarian carcinoma (Xiao et al., 2000; 2001; Xu et al., 2001; Sutphen et al., 2004; Murph et al., 2007; Pineiro et al., 2011; Pineiro and Falasca, 2012). To test our hypothesis, that ovarian cancer cells secrete LPI, and thus promote tumour angiogenesis in vivo via an LPI/GPR55-dependent mechanism; conditioned medium from the human ovarian cancer cell lines OVCAR-3, OVCAR-5 and COV-362 was analysed for its LPI levels and in the CAM angiogenesis model. LC-MS/MS revealed that OVCAR-3, OVCAR-5 and COV-362 cells produced significant but quite different amounts of LPI (Figure 1A). Within 3 days, conditioned medium from OVCAR-3, OVCAR-5 and COV-362 strongly induced angiogenesis in vivo to a similar extent (90–100% increase), compared with unconditioned medium (Figure 1B). Selective inhibition of the LPI receptor GPR55 with CID16020046 (20 μM) effectively blocked ovarian cancer-induced angiogenesis of all tested cell lines (Figure 1B). Together, these results suggest that LPI produced by ovarian cancer cells induces angiogenesis in a GPR55-dependent manner.

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