scholarly journals Proteomic Characterization of Angiogenic Endothelial Cells Stimulated with Cancer Cell-Conditioned Medium

2007 ◽  
Vol 30 (12) ◽  
pp. 2300-2307 ◽  
Author(s):  
Yasufumi Katanasaka ◽  
Tomohiro Asai ◽  
Hirotaka Naitou ◽  
Norio Ohashi ◽  
Naoto Oku
Keyword(s):  
Endothelial Cells ◽  
Conditioned Medium ◽  
Cancer Cell ◽  
Cell Conditioned Medium

NIDOGEN1 as a novel regulator of endothelial control over breast cancer invasion and metastasis.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e23005-e23005
Author(s):  
Martin Buess ◽  
Daniela A. Ferraro ◽  
Francesca Patella ◽  
Sara Zanivan ◽  
Gerhard Christofori
Keyword(s):  
Breast Cancer ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Conditioned Medium ◽  
Cancer Cell ◽  
Cancer Biology ◽  
Invasion And Metastasis ◽  
Stat3 Pathway ◽  
Endothelial Control ◽  
And Migration

e23005 Background: The microenvironment is a central regulator of cancer biology. While the contribution of fibroblasts has been largely studied, the role of endothelial cells as regulators of cancer cell behavior is still poorly understood. As in a diverse spectrum of physiological processes in normal tissue, endothelial cells may exert a similar regulatory control in cancer progression and metastasis. Methods: To characterize the functional effects of endothelial-cancer interaction we focused on an in vitro co-culture model. Results: Co-culturing human umbilical venous endothelial cells (HUVEC) with SKBR-3 breast cancer cells induced morphological changes with epithelial-mesenchymal transition traits (EMT) and a significantly increased migratory and invasive potential. This activity leading to an elongated phenotype, expression of mesenchymal markers and pro-migratory gene sets in SKBR-3 was contained in HUVEC conditioned medium. The pro-migratory effect on SKBR-3 was significantly more pronounced when the supernatant was obtained from a sub-confluent and highly proliferative endothelial cell culture than from a confluent and resting endothelial cell layer. To identify the secreted regulatory molecules, we analyzed the supernatant of sub-confluent and confluent endothelial cells by quantitative MS proteomics (SILAC analysis). Eight candidate proteins significantly more secreted in conditioned medium from confluent HUVEC represented potential inhibitors of migration. Among them NIDOGEN1 was found to be necessary and sufficient for the inhibition of EMT and migration in SKBR-3. Stimulation of SKBR-3 with supernatant from sub-confluent HUVEC increased p-STAT3 levels in SKBR-3. Silencing nidogen1 in confluent HUVEC re-activated phosphorylation of STAT3 indicating that NIDOGEN1 inhibits the promigratory STAT3 pathway. The STAT3 pathway and migration were also inhibited by overexpression of nidogen1 in MDA-MB-231 LM2 cells.When injected in the mammary fat pad of nude mice these cells formed significantly less lung metastases than controls (p < 0.01). Conclusions: We identified NIDOGEN1 as a novel regulator of endothelial control over cancer cell migration, invasion and metastasis.

Isolation and characterization of a neu protein-specific activating factor from human ATL-2 cell conditioned medium

1991 ◽  
Vol 179 (3) ◽  
pp. 1536-1542 ◽  
Author(s):  
James G. Davis ◽  
Junji Hamuro ◽  
Caroline Y. Shim ◽  
Arabinda Samanta ◽  
Mark I. Greene ◽  
...  
Keyword(s):  
Conditioned Medium ◽  
Isolation And Characterization ◽  
Cell Conditioned Medium

Endothelium-dependent ANF secretion in vitro

1992 ◽  
Vol 263 (4) ◽  
pp. H1071-H1077 ◽  
Author(s):  
R. A. Lew ◽  
A. J. Baertschi
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Conditioned Medium ◽  
Superoxide Anion ◽  
Heat Stable ◽  
Guanylate Cyclase Activator ◽  
Stimulatory Factor ◽  
Cell Conditioned Medium ◽  
Stimulation Of

Coculture of endothelial cells with atrial cells (R. A. Lew and A. J. Baertschi. Biochem. Biophys. Res. Commun. 163: 701-709, 1989) increased atrial natriuretic factor (ANF) release to 205 +/- 15% (n = 33 experiments) of basal secretion (2.02 +/- 0.33 ng/ml). Stimulation of ANF release by endothelial cells was significantly reduced (P < 0.05) by addition of the calcium channel antagonist nicardipine (Nic, 100 nM; by 69 +/- 4%), the guanylate cyclase activator sodium nitroprusside (SNP, 1 microM; by 97 +/- 27%), or acetylcholine (ACh, 10 microM; by 55 +/- 13%). Endothelial cell-conditioned medium elicited a 62 +/- 10% (n = 10) increase in ANF release. Rat and porcine endothelin (0.1-100 nM) each elicited a dose-dependent increase in ANF release [up to 84 +/- 14% (n = 18) over baseline]. The activity of conditioned medium was not affected by heat or trypsin treatment, but was significantly reduced by addition of Nic or SNP and was attenuated by ACh. Stimulation of ANF by 1 nM synthetic rat or porcine endothelin was also unaffected by heat or trypsin but was significantly reduced by Nic, SNP, and ACh. Addition of endothelin-specific antiserum abolished the ANF stimulatory activity of endothelial cell-conditioned medium. Neither inhibition of superoxide anion by superoxide dismutase nor inhibition of endothelium-derived nitric oxide production by NG-monomethyl-L-arginine affected the ANF release from coculture. Thus endothelial cells release a heat-stable, diffusible ANF stimulatory factor, which is not endothelium-derived relaxing factor or superoxide anion but is biologically and immunologically similar to endothelin.

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