12 research outputs found

    The Hippo pathway is responsible for S100A7 induction in well differentiated cervical and pharyngeal SCC cells.

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    <p>(a, b) Overexpression of LATS1 in normal attached HCC94 (a, left), FaDu (a, right), SiHa (b, left) and H226 (b, right) cells. Anti-Flag tag antibody was used to judge the transfection efficiency. HCC94-WT was used as the positive control. β-actin was used as a loading control (c, d) Depletion of LATS1 using siRNA attenuates S100A7 in suspended or dense HCC94 (c) and FaDu (d) cells. siLATS1+S48h (or D48h) indicates that cells are cultured in suspension (or dense) for 48 h after silencing of LATS1. (e, f) Silencing MST1 in HCC94 (e) and FaDu (f) cells cultured in suspension or dense. siMST1+S48h (or D48h) indicates that cells are cultured in suspension (or dense) for 48 h after losing of MST1. GAPDH was used as a loading control.</p

    TEAD1 mediates YAP-dependent S100A7 expression.

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    <p>(a) Western blot analyses for S100A7 expression after TEAD1 silencing in normal attached HCC94 cells. (b) The mRNA expression of <i>TEAD1</i>, <i>CTGF</i>, <i>CYR61</i> and <i>S100A7</i> is examined using qRT-PCR. H: HCC94 cells. Error bar, SD of three different experiments. <i>*P<0</i>.<i>05</i>, **<i>P</i><0.01; <i>t-test</i>. (c, d) Overexpression of YAP-WT and mutant activated YAP-S94A in normal attached HCC94 (c) and FaDu (d) cells. Anti-Flag tag antibody was used to judge the transfection efficiency. β-actin was used as a loading control.</p

    The precautionary principle and genetically modified organisms: a bone of contention between European institutions and member states

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    Theoretical thesis.Bibliography: pages 75-86.Introduction -- Chapter I. The precautionary principle -- Chapter II. The precautionary principle and genetically modified organisms under the European legal framework -- Chapter III. The precautionary principle and genetically modified organisms : analysis of three seminal cases of the European Court of Justice (2003-2011) -- Chapter IV. A high level of protection for health and the environment vs free circluation of GMOs on the European market : interests in conflict -- Bibliography.This dissertation examines how the Precautionary Principle, as an internationally recognised concept enshrined in a range of legal instruments, has been applied to provide a mechanism for protection of the environment and health in response to the introduction of Genetically Modified Organisms (GMOs) in Europe. It examines how the European Court of Justice substantively handled the risk assessment phase across three seminal cases between 2003 and 2011 in which Member States had failed in their attempt to trigger the Precautionary Principle in order to uphold a ban or suspension of the cultivation or sale of products derived from GMOs in their territory. The analysis of these judgements suggests that the Court has applied a narrow approach to the evidence provided by national governments during the risk assessment stage, and has thereby limited the potential for precautionary measures by Member States to be upheld by the Court. This outcome reflects a ‘weak’ application of the Precautionary Principle by the Court in contrast with the ‘strong’ interpretation implied by the European legal and policy framework and objections of Member States.Mode of access: World wide web1 online resource (86 pages

    The Characteristics and Function of S100A7 Induction in Squamous Cell Carcinoma: Heterogeneity, Promotion of Cell Proliferation and Suppression of Differentiation

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    <div><p>S100A7 is highly expressed in squamous cell carcinomas (SCC) and is related to the terminal differentiation of keratinocytes. However, its characteristic and function in SCC is not very known. In this present study, we used immunohistochemistry to examine the expression of S100A7 in 452 SCC specimens, including the lung, esophagus, oral cavity, skin, cervix, bladder, and three SCC cell lines. We found that S100A7-positive staining showed significant heterogeneity in six types of SCC specimen and three SCC cell lines. Further examination found that S100A7-positive cells and its expression at mRNA and protein levels could be induced in HCC94, FaDu, and A-431 cells both in vitro and in vivo using immunohistochemistry, real-time PCR, and Western blotting. Notably, the upregulation of squamous differentiation markers, including keratin-4, keratin-13, TG-1, and involucrin, also accompanied S100A7 induction, and a similar staining pattern of S100A7 and keratin-13 was found in HCC94 cells both in vitro and in vivo. Further study revealed that the overexpression of S100A7 significantly increased proliferation and inhibited squamous differentiation in A-431 cells both in vitro and in vivo. Conversely, silencing S100A7 inhibited cell growth and survival and increased the expression of keratin-4, keratin-13, TG-1, and involucrin in HCC94 cells. Therefore, these results demonstrate that S100A7 displays heterogeneous and inducible characteristic in SCC and also provide novel evidence that S100A7 acts as a dual regulator in promoting proliferation and suppressing squamous differentiation of SCC.</p></div

    Squamous differentiation markers were also induced by suspension and confluent treatment.

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    <p>The expression of keratin-1, keratin-13, keratin-4, TG-1 and involucrin was determined by real-time PCR for the FaDu (A); A-431(B) and HCC94 (C, F) cells after suspension or confluent culture. The protein level of keratin-13 was confirmed by Western blotting for the FaDu (D) and HCC94 (E) cells. S48h: suspension 48h. N: pre-suspension. C6: cells were cultured at confluence for six days. C6-R3P: cells were cultured at confluence for six days and then reseeded at sub-confluence for 3 cell passages. *, p<0.05, and **, p<0.01 versus the S100A7 experiential groups and the control groups. The results represent the average values from three independent experiments.</p

    The expression of S100A7 in SCC tissues.

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    <p>SCC and normal tissues were examined by immunohistochemistry with specific anti-S100A7 antibody in lung (A, M); esophagus (B, N); cervix (C, O); bladder (D, P); oral cavity (E, Q); skin (F, R). The corresponding types of SCC tissues were also examined by immunohistochemistry with nonspecific IgG (G-L). Arrowheads indicate the positive staining of S100A7 and the asterisks indicate the keratinizing areas.</p

    S100A7 promotes squamous carcinoma cell growth and inhibits squamous cells differentiation both in vitro and in vivo.

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    <p>The proliferation and the survival rate of SCC cells were evaluated by MTT (A, B, C) and XTT (D, E). The expression of differentiation markers was determined by real-time PCR (F-L). The expressions of S100A7 and keratin-13 after overexpression or knockdown S100A7 in SCC cells were confirmed by Western blotting (M, N, O). ‘NC’ stands for mock-transfected cells. The volumes of tumor xenografts derived from the stable-overexpressing S100A7-A431 cells and the control cells (P). The expression of differentiation markers in xenografts was examined by real-time PCR (Q-S). The stable-overexpressing S100A7 in A-431 cells was confirmed by Western blotting (T). *, p<0.05, and **, p<0.01 versus the S100A7 experiential groups and the control groups. The results represent the average values from six independent experiments.</p

    The induction of S100A7 expression in HCC94 cells in confluent culture.

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    <p>S100A7 expression was examined by immunohistochemistry (A, C, E), real-time PCR (G) and Western blotting (H). Cells were also examined by immunohistochemistry with nonspecific IgG (B, D, F).*, p<0.05, and **, p<0.01 versus the experimental groups and the pre-confluent groups. The results represent the average values from three independent experiments. N: pre-confluence; C6: cells were cultured at confluence for 6 days; C6-R3P: cells that were cultured at confluence for six days and then reseeded at sub-confluence for 3 cell passages.</p

    The induction of S100A7 expression in FaDu, A-431, and HCC94 cells in suspension culture.

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    <p>The expression of S100A7 was determined by real-time PCR (A-C) and Western blotting (D-F). FaDu, HCC94 and A-431cells in suspension for two days were reattached for 12h and examined using immunohistochemistry with specific anti-S100A7 antibody (G-I) and nonspecific IgG (J-L). S48h: suspension 48h. N: pre-suspension.*, p<0.05, and **, p<0.01 versus the suspension groups and the pre-suspension groups. The results represent the average values from three independent experiments.</p
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