S100A10 regulates plasminogen-dependent macrophage invasion

Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada.
Blood (Impact Factor: 10.45). 08/2010; 116(7):1136-46. DOI: 10.1182/blood-2010-01-264754
Source: PubMed


The plasminogen activation system plays an integral role in the migration of macrophages in response to an inflammatory stimulus, and the binding of plasminogen to its cell-surface receptor initiates this process. Although previous studies from our laboratory have shown the importance of the plasminogen receptor S100A10 in cancer cell plasmin production, the potential role of this protein in macrophage migration has not been investigated. Using thioglycollate to induce a peritoneal inflammatory response, we demonstrate, for the first time, that compared with wild-type (WT) mice, macrophage migration across the peritoneal membrane into the peritoneal cavity in S100A10-deficient (S100A10(-/-)) mice was decreased by up to 53% at 24, 48, and 72 hours. Furthermore, the number of S100A10-deficient macrophages that infiltrated Matrigel plugs was reduced by 8-fold compared with their WT counterpart in vivo. Compared with WT macrophages, macrophages from S100A10(-/-) mice demonstrated a 50% reduction in plasmin-dependent invasion across a Matrigel barrier and a 45% reduction in plasmin generation in vitro. This loss in plasmin-dependent invasion was in part the result of a decreased generation of plasmin and a decreased activation of pro-MMP-9 by S100A10-deficient macrophages. This study establishes a direct involvement of S100A10 in macrophage recruitment in response to inflammatory stimuli.

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    • "It is also reported that ANXA2 may play a key role in the retinal neovascularization. With its partner S100A10 (p11), ANXA2 is able to form a heterotetrameric complex [3– 10], which binds both plasminogen and its activator (tissue plasminogen activator, tPA) to accelerate the generation of plasmin [11] [12] [13] [14] [15] [16] [17]. The isolated endothelial cells from the mice deficient in annexin A2 (ANXA2 −/− ) are unable to support tPA-dependent plasminogen activation in vitro [18] [19] and the ANXA2 −/− mice exhibit reduced angiogenesis in growth factor-stimulated assays in adulthood. "
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    ABSTRACT: Objective: To study the effects of inhibited Annexin A2 (ANXA2) on human umbilical vein endothelial cells (HUVECs) in vitro. Methods: Short hairpin RNA (shRNA) targeting ANXA2 was designed and cloned into double marked lentivirial vector GV248 for RNAi to generate the recombinant expression plasmids, which were stably transfected into HUVECs. The protein and mRNA expression levels of ANXA2 were analyzed by western blotting and real-time polymerase chain reaction, respectively. Cell proliferation (cell counting kit-8 assay), apoptosis (flow cytometry analysis), the expression (western blotting) and the activity of caspases (enzyme-linked immunosorbent assay) were used to assess the effects of silencing ANXA2 on HUVECs in vitro. Results: The plasmids to express ANXA2-specific shRNA were constructed and were infected into HUVEC resulting in the stably transfected experimental (ANXA2-shRNA), control (control-shRNA) and mock (no plasmid) cell lines, which were verified with western blot and real-time PCR. HUVEC/ANXA2-shRNA showed an inhibition rate 91.89% of ANXA2 expression compared to the mock HUVEC. ANXA2 silencing cell strain obviously presented a lower cell proliferation activity compared to the control and mock HUVECs, with an inhibition rate 82.35% on day 7 in vitro. FACS analysis indicated that the HUVEC/ANXA2-shRNA cells undergoing apoptosis increased by 102.61% compared to the mock HUVECs (P < 0.01). Moreover, the activity levels of caspase-3, caspase-8 and caspase-9 in HUVEC/ANXA2-shRNA cells were increased and the activated cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9 were upregulated evidently compared with that of the control and mock HUVECs by 56.29%, 89.59% and 144.58% (P < 0.01). Conclusions: shRNA-mediated silencing of ANXA2 could not only be able to suppress HUVECs proliferation but to upregulate the enzyme activity of caspases, which bring to an increase of cell apoptosis. This work suggested that ANXA2 may represent a useful target of future molecular therapies.
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    • "Given the detailed knowledge of the binding interactions between the N-terminus of AnxA2 and S100A10, various groups have reported the use of peptides based upon this N-terminus to perform competition experiments with the aim of disrupting the endogenous complex of the two proteins and understanding its functions. An isolated acetylated synthetic peptide comprising residues 1–14 of AnxA2 can disrupt a preformed complex between S100A10 and a labelled annexin 1–14 peptide (O'Connell et al., 2010). Furthermore, the same peptide also disrupts a preformed full-length (S100A10-AnxA2)2 complex (Konig et al., 1998). "
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    • "In mice, S100A4 -/ -animals showed reduced joint inflammation , impaired bone resorption and regulation of osteoclast, increased cartilage and bone destruction, and decreased expression of MMPs (Bian, 2011; Erlandsson et al., 2013). S100A10 expression decreased the production of TNFa, IL- 1b, and IL-10 in chondrocytes (O'Connell et al., 2010; Song et al., 2012). The S100A11 induces chondrocyte hypertrophy and matrix catabolism (Yammani, 2012). "
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