A2PP reduces <i>M</i>. <i>hyorhinis</i> infection.

<p>Cell ELISA analysis of p37 (OD 490 nm) of p37 protein in AGS and BGC823 cells infected with 10⁵ CCU (color changing units)/ml of <i>M</i>. <i>hyorhinis</i> and treated with A2PP or ConP for 24 hr. <i>M</i>. <i>hy</i>, <i>M</i>. <i>hyorhinis</i>. Mean ± SD from 3 experiments with triplicate for each sample. (B) Quantitative PCR (qPCR) analysis of <i>p37</i> in AGS and BGC823 cells infected and treated as in (A). Mean ± SD from 3 experiments with triplicate for each sample. (C) Western blotting of p37, p-EGFR, EGFR, p-ANXA2 and ANXA2 from AGS and BGC823 cells treated as in (A). (D) Quantification of p37 protein levels of in (C). Levels of p37 were normalized to those of GAPDH. Mean ± SD from 3 independent experiments. (E) Quantification of p-EGFR and p-ANXA2 levels in (C). Levels of p-EGFR or p-ANXA2 were normalized to those of EGFR or ANXA2. Mean ± SD from 3 independent experiments. **, P < 0.01; **, P < 0.001; n.s, no significance.</p

A2PP binds to p37 of <i>M</i>. <i>hyorhinis</i> and has minimal effect on the proliferation of gastric cancer cell.

<p>(A) Schematic diagram of amino acid sequence of N-terminal of ANXA2 polypeptide (A2PP). (B) Solid-phase binding assay. OD490 of 15 nM biotin-A2PP to GST was set as 1 and relative bindings were caculated. Mean ± SD of three independent assays with triplicate samples. ***, P < 0.001. (C) Streptavidin pull-down assays identified Biotin-A2PP as a GST-p37 binding polypeptide. (D) Cell morphology of AGS and BGC823 following indicated concentrations of A2PP treatment for 24 hr and 48 hr. (E) Proliferation of gastric cancer cell lines (AGS and BGC823) treated with increasing concentration of A2PP for 72 hr. Mean ± SD from three experiments with triplicate samples.</p

A2PP has minimal effects on EGFR-ANXA2 signaling, migration of gastric cancer cells, or the localization of ANXA2.

<p>(A) Western blotting of phospho-EGFR, phospho-ANXA2, EGFR, and ANXA2 from AGS and BGC823 cells treated with A2PP for 24 hr. GAPDH was used as loading control. (B) Representative images of migration of AGS and BGC823 cells treated with A2PP for 24 hr. Scale bars, 200 μm. (C) Statistical summary of migration assay. Mean ± SD from three experiments with triplicate samples. ns, no significance. (D) Immunofluorescence of ANXA2 localization (red) in AGS and BGC823 cells treated with A2PP for 24 hr. Scale bars, 5 μm.</p

A2PP inhibits <i>M</i>. <i>hyorhinis</i> infection.

<p>(A) Western blotting of p37 from AGS and BGC823 cells infected with 10⁵ CCU/ml of <i>M</i>. <i>hyorhinis</i> and treated with A2PP (20 μM), CIP (4 μg/ml), MYCO I (5 μg/ml), or MYCO II (10 μg/ml) for 24 hr. Mean ± SD from 3 independent experiments. (B) qPCR analysis of <i>p37</i> in AGS and BGC823 cells treated as in (A). Mean ± SD from 3 experiments with triplicate for each sample. (C) Western blotting of p37 from AGS cells infected with 10⁵ CCU/ml of <i>M</i>. <i>hyorhinis</i> and treated with A2PP in the process of cell passages from P₁ to P₃. Mean ± SD from 3 independent experiments. (D) qPCR analysis of <i>p37</i> in AGS cells infected and treated as in (C). Mean ± SD from 3 experiments with triplicate samples. **, P < 0.01; ***, P < 0.001; ns, no significance.</p

A2PP have a less cytotoxicity.

<p>(A) Proliferation of AGS and BGC823 treated with A2PP (20 μM), CIP (4 μg/ml), MYCO I (5 μg/ml), or MYCO II (10 μg/ml) for 72 hr. Mean ± SD from 4 independent experiments with triplicate samples. (B) RT-PCR analysis of <i>ATF5</i>, <i>DDIT3</i>, <i>CEBPB</i> in AGS and BGC823 cells treated with A2PP (10 μM), CIP (4 μg/ml), MYCO I (5 μg/ml), MYCO II (10 μg/ml) for 24 hr. Mean ± SD from 3 experiments with triplicate samples.</p

A2PP suppresses migration of gastric cancer cell induced by <i>M</i>. <i>hyorhinis</i> infection.

<p>(A) Migration of 10⁵ CCU/ml of <i>M</i>. <i>hyorhinis</i>-infected AGS and BGC823 cells treated with indicated peptides for 24 hr. (B) Summary of migration assays (n = 3). Mean ± SD from 3 independent experiments with triplicate samples. ***, P<0.001.</p

Specific sequence of A2PP decreases <i>M</i>. <i>hyorhinis</i> infection.

<p>Schematic diagram of two truncated forms of A2PP. (B) qPCR analysis of <i>p37</i> in AGS and BGC823 cells infected with 10⁵ CCU/ml of <i>M</i>. <i>hyorhinis</i> and treated with 20 μM indicated peptides for 24 hr. Mean ± SD from 3 experiments with triplicate for each sample. (C) Western blotting of p37 from AGS and BGC823 cells infected and treated as in (B). Mean ± SD from 3 independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.</p

Overall and group-specific summary statistics for <i>TNF-a</i> 308, <i>TNF-a</i> 238 in colorectal cancer.

<p>CI: confidence interval; OR: odds ratio; <i>TNF-a</i>: tumor-necrosis factor-a.</p

Additional file 1: of Extracellular gamma-synuclein promotes tumor cell motility by activating β1 integrin-focal adhesion kinase signaling pathway and increasing matrix metalloproteinase-24, -2 protein secretion

Table S1. Detailed information of antibodies used in the study. Table S2. Clinicopathologic characteristics of patients (n=250). Table S3. Small interfering RNA sequences for β1 and FAK. Figure S1. Effect of small interfering RNA sequence on β1 protein expression. Figure S2. β1 integrin and FAK mediate SNCG-promoted tumor cell migration. Figure S3. Up-regulation of phospho-FAK induced by SNCG is blocked by β1 integrin knockdown, but has no effect on Src or Erk phosphorylation. Figure S4. Exogenously added SNCG promotes MMP-24 and MMP-2 secretion from colorectal cancer cells. (DOC 1280 kb

The association between <i>TNF</i>-a 238 polymorphism and risk of colorectal cancer.

<p>A: AA vs. GG, Forest plot; B: AG vs. GG, Forest plot; ORs, odds ratios.</p