Ovarian cancer ascites-derived vitronectin and fibronectin: Combined purification, molecular features and effects on cell response

Carduner, Ludovic; Agniel, Rémy; Kellouche, Sabrina; Picot, Cědric R.; Blanc-Fournier, Cécile; Leroy-Dudal, Johanne; Carreiras, Franck

doi:10.1016/j.bbagen.2013.06.023

Cited by 21 publications

(22 citation statements)

References 42 publications

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“…FLRT2‐ECD precipitates immunoblotted with Fn anti‐serum revealed the presence of a strong band of approximately 250–300 kDa (lane 4, Fig. ‐I), corresponding to the previously documented possible molecular weights of Fn (Carduner et al, ; Lemańska‐Perek et al, ). Blotting of the stripped membrane with FLRT2‐ECD Ab demonstrated the presence of an approximate 80 kDa band in Fn‐immunoprecipitates (lane 5, Fig.…”

Section: Resultssupporting

confidence: 72%

FLRT2 Interacts With Fibronectin in the ATDC5 Chondroprogenitor Cells

Arudchelvan

Gong

2014

Journal Cellular Physiology

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Expression studies have implicated FLRT2 in cranial neural crest cell migration and prechondrogenic cell condensation during craniofacial skeletogenesis. We aimed to determine whether FLRT2 was involved in mediating cell-matrix interactions in the ATDC5 chondroprogenitor cell line. Immunolocalization experiments of ATDC5 cells revealed that FLRT2 was present on the cell membrane as well as extracellularly, where it colocalized with Fibronectin (Fn). After cell extraction of the matrix, FLRT2 was identified in the ATDC5-derived extracellular matrix (ECM) and was further found to be associated with Fn-coated beads in cell cultures. Blockage of Fn fibril formation via a blocking peptide resulted in a concomitant decrease in extracellular FLRT2 accumulation. Over a 7-day period following the replenishment of the Fn blocking peptide to the cultures, there was a partial rebound in Fn fibril formation that was accompanied by a concomitant reappearance of FLRT2 co-expression. Co-immunoprecipitation confirmed that FLRT2 and Fn interacted, either directly or indirectly. Immunoprecipitation and Western blot analyses with antibodies recognizing epitopes located on the extra- and intracellular domains of FLRT2 further revealed the presence of different sized bands, suggesting that FLRT2 may exist in both membrane-bound and shed forms. Our data therefore provide evidence that FLRT2 and/or its cleavage products may be cooperating with Fn and other ECM proteins to regulate critical cellular events. Further studies will be necessary in delineate more precisely the roles of FLRT2 in mediating cell- and cell-matrix interactions during normal development.

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Section: Resultssupporting

confidence: 72%

FLRT2 Interacts With Fibronectin in the ATDC5 Chondroprogenitor Cells

Arudchelvan

Gong

2014

Journal Cellular Physiology

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“…For example, vitronectin is an important factor in the progression of ovarian cancer, and has been previously assayed in ascites fluid [27]. More importantly, cleavage of vitronectin and fibronectin into small fragments (mediated by MMP-2) is a key initiator of metastasis in ovarian cancer [28].…”

Section: Resultsmentioning

confidence: 99%

Deciphering the ovarian cancer ascites fluid peptidome

et al. 2014

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BackgroundConventional proteomic approaches have thus far been unable to identify novel serum biomarkers for ovarian cancer that are more sensitive and specific than the current clinically used marker, CA-125. Because endogenous peptides are smaller and may enter the circulation more easily than proteins, a focus on the low-molecular-weight region may reveal novel biomarkers with enhanced sensitivity and specificity. In this study, we deciphered the peptidome of ascites fluid from 3 ovarian cancer patients and 3 benign individuals (ascites fluid from patients with liver cirrhosis).ResultsFollowing ultrafiltration of the ascites fluids to remove larger proteins, each filtrate was subjected to solid phase extraction and fractionated using strong cation exchange chromatography. The resultant fractions were analyzed using an Orbitrap mass spectrometer. We identified over 2000 unique endogenous peptides derived from 259 proteins. We then catalogued over 777 peptides that were found only in ovarian cancer ascites. Our list of peptides found in ovarian cancer specimens includes fragments derived from the proteins vitronectin, transketolase and haptoglobin.ConclusionsPeptidomics may uncover previously undiscovered disease-specific endogenous peptides that warrant further investigation as biomarkers for ovarian cancer.

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“…It has been demonstrated to play an important role in the adhesion of cancer cells to integrins. Specifically, vitronectin has been shown to promote invasiveness of ovarian cancer cells and their spread in the peritoneum [20, 21]. The mechanisms responsible are related to MMP-2-mediated cleavage of vitronectin and fibronectin to generate fragments that interact with integrin receptors α 5 β 1 and α v β 3 , which promote the adhesion of ovarian cancer cells lining the peritoneal cavity [22].…”

Section: Discussionmentioning

confidence: 99%

Evidence that vitronectin is a potent migration-enhancing factor for cancer cells chaperoned by fibrinogen: a novel view of the metastasis of cancer cells to low-fibrinogen lymphatics and body cavities

et al. 2016

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Diluted (1%) plasma induces migration of malignant cell lines much more strongly than potent pro-metastatic factors. To characterize the factor(s) present in diluted plasma responsible for this phenomenon we performed i) heat inactivation, ii) dialysis, iii) proteinase K treatment, and iv) molecular size filtration studies. We found that this remarkable pro-migratory activity of diluted normal plasma is associated with a ~50–100-kD protein that interacts with GαI protein-coupled receptors and activates p42/44 MAPK and AKT signaling in target cells. Since this pro-migratory activity of 1% plasma decreases at higher plasma concentrations (> 20%), but is retained in serum, we hypothesized that fibrinogen may be involved as a chaperone of the protein(s). To identify the pro-migratory protein(s) present in diluted plasma and fibrinogen-depleted serum, we performed gel filtration and hydrophobic interaction chromatography followed by mass spectrometry analysis. We identified several putative protein candidates that were further tested in in vitro experiments. We found that this pro-migratory factor chaperoned by fibrinogen is vitronectin, which activates uPAR, and that this effect can be inhibited by fibrinogen. These results provide a novel mechanism for the metastasis of cancer cells to lymphatics and body cavities, in which the concentration of fibrinogen is low, and thus suggests that free vitronectin stimulates migration of tumor cells.

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Ovarian cancer ascites-derived vitronectin and fibronectin: Combined purification, molecular features and effects on cell response

Cited by 21 publications

References 42 publications

FLRT2 Interacts With Fibronectin in the ATDC5 Chondroprogenitor Cells

FLRT2 Interacts With Fibronectin in the ATDC5 Chondroprogenitor Cells

Deciphering the ovarian cancer ascites fluid peptidome

Evidence that vitronectin is a potent migration-enhancing factor for cancer cells chaperoned by fibrinogen: a novel view of the metastasis of cancer cells to low-fibrinogen lymphatics and body cavities

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