A magyar közlekedéspolitika stratégiai környezeti vizsgálata

A stratégiai környezeti vizsgálatok nemzetközi szakirodalmából származó tapasztalatokkal összhangban a közlekedéspolitika vizsgálatához a stratégiai környezeti vizsgálat szélesebb értelmezésének elfogadásával egy általános fenntarthatósági vizsgálat elvégzésére irányuló módszer kidolgozását kezdeményeztük. Ennek keretében a közlekedéspolitika cél- és intézkedési rendszerét elemeztük a célok belső és külső konzisztenciája szempontjából, illetve az egyes célokat összevetettük a fenntarthatóság kritériumaival

Syndecan-1 Enhances Proliferation, Migration and Metastasis of HT-1080 Cells in Cooperation with Syndecan-2

Syndecans are transmembrane heparan sulphate proteoglycans. Their role in the development of the malignant phenotype is ambiguous and depends upon the particular type of cancer. Nevertheless, syndecans are promising targets in cancer therapy, and it is important to elucidate the mechanisms controlling their various cellular effects. According to earlier studies, both syndecan-1 and syndecan-2 promote malignancy of HT-1080 human fibrosarcoma cells, by increasing the proliferation rate and the metastatic potential and migratory ability, respectively. To better understand their tumour promoter role in this cell line, syndecan expression levels were modulated in HT-1080 cells and the growth rate, chemotaxis and invasion capacity were studied. For in vivo testing, syndecan-1 overexpressing cells were also inoculated into mice. Overexpression of full length or truncated syndecan-1 lacking the entire ectodomain but containing the four juxtamembrane amino acids promoted proliferation and chemotaxis. These effects were accompanied by a marked increase in syndecan-2 protein expression. The pro-migratory and pro-proliferative effects of truncated syndecan-1 were not observable when syndecan-2 was silenced. Antisense silencing of syndecan-2, but not that of syndecan-1, inhibited cell migration. In vivo, both full length and truncated syndecan-1 increased tumour growth and metastatic rate. Based on our in vitro results, we conclude that the tumour promoter role of syndecan-1 observed in HT-1080 cells is independent of its ectodomain; however, in vivo the presence of the ectodomain further increases tumour proliferation. The enhanced migratory ability induced by syndecan-1 overexpression is mediated by syndecan-2. Overexpression of syndecan-1 also leads to activation of IGF1R and increased expression of Ets-1. These changes were not evident when syndecan-2 was overexpressed. These findings suggest the involvement of IGF1R and Ets-1 in the induction of syndecan-2 synthesis and stimulation of proliferation by syndecan-1. This is the first report demonstrating that syndecan-1 enhances malignancy of a mesenchymal tumour cell line, via induction of syndecan-2 expression

Dynamic Interplay in Tumor Ecosystems: Communication between Hepatoma Cells and Fibroblasts

Tumors are intricate ecosystems where cancer cells and non-malignant stromal cells, including cancer-associated fibroblasts (CAFs), engage in complex communication. In this study, we investigated the interaction between poorly (HLE) and well-differentiated (HuH7) hepatoma cells and LX2 fibroblasts. We explored various communication channels, including soluble factors, metabolites, extracellular vesicles (EVs), and miRNAs. Co-culture with HLE cells induced LX2 to produce higher levels of laminin β1, type IV collagen, and CD44, with pronounced syndecan-1 shedding. Conversely, in HuH7/LX2 co-culture, fibronectin, thrombospondin-1, type IV collagen, and cell surface syndecan-1 were dominant matrix components. Integrins α6β4 and α6β1 were upregulated in HLE, while α5β1 and αVβ1 were increased in HuH7. HLE-stimulated LX2 produced excess MMP-2 and 9, whereas HuH7-stimulated LX2 produced excess MMP-1. LX2 activated MAPK and Wnt signaling in hepatoma cells, and conversely, hepatoma-derived EVs upregulated MAPK and Wnt in LX2 cells. LX2-derived EVs induced over tenfold upregulation of SPOCK1/testican-1 in hepatoma EV cargo. We also identified liver cancer-specific miRNAs in hepatoma EVs, with potential implications for early diagnosis. In summary, our study reveals tumor type-dependent communication between hepatoma cells and fibroblasts, shedding light on potential implications for tumor progression. However, the clinical relevance of liver cancer-specific miRNAs requires further investigation

List of recognition sequences targeted by silencer plasmids used.

<p>S1miRNA-a and -b are specific for syndecan-1, S2miRNA-a and -b target syndecan-2.</p

Effects of FullEGFP and 78Sig on the proliferation and chemotactic migration of HT-1080 cells.

<p>(A) Doubling times were calculated from the log phase of growth curves (B) obtained from the SRB colorimetric assay. Values are expressed as mean±s.d. (n = 8). (C) CDK2, phospho-retinoblastoma (at T373 position) and GAPDH immunoblots of HT-1080 cells transfected with EGFP, FullEGFP or 78Sig. (D) Representative fields of cyclin E immunocytochemistry (red) on cultured HT-1080 cells transfected with EGFP, FullEGFP or 78Sig. Scale bar: 20 µm. (E) Relative amounts of migrated cells toward ECM proteins in a Boyden chamber after transfection with EGFP, FullEGFP or 78Sig. Values are shown as mean±s.d. (n = 5), *p<0.05 and **p<0.01. (F) Syndecan-2 and GAPDH immunoblots of cultured HT-1080 transfectants. Sdc-2 stands for syndecan-2 transfection. (G) Results of flow cytometry after immunofluorescent staining of stable transfectants using the ZMD.308 antibody for syndecan-2. (H) Immunofluorescent staining of methanol-fixed cells by the syndecan-2 specific antibody L-18 (red). Identical exposure times and background corrections were applied. Scale bar: 50 µm. For (E and G) nuclei were counterstained with DAPI (blue).</p

The area percentage of lung metastases.

<p>Mice were sacrificed on the 50th day after injection of HT-1080 cells into their foot pads expressing EGFP, FullEGFP or 78Sig. Area percentage referring to the area fractions of lung metastases were calculated by morphometrical analysis of hematoxylin-eosin stained sections of the lungs in 5 random planes.</p

Candidates in the molecular mechanism of the cooperation between syndecan-1 and syndecan-2.

<p>(A) Results of pRTK array. Relative extents of IGF1R and Axl phosphorylation in HT-1080 transfectants. Values are expressed as mean±s.d. *p<0.05 versus control EGFP cells (n = 3). (B and C) Representative immunoblots from HT-1080 cells stably expressing EGFP, FullEGFP, 78Sig or Sdc-2. Antibodies used are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039474#pone.0039474.s002" target="_blank">Table S1</a>. (D) Results of densitometry of western blots. Values are expressed as mean±s.d. *p<0.05 by <i>t</i>-test.</p