BRMS1staining and clinicopathological characteristics of 192 glioma patients.

<p>*χ² test.</p

Overexpression of BRMS1 inhibits glioma cells adhesion ability.

<p><b>A, B</b> Cell attachment assay after BRMS1 overexpression in U251 and U87 cells. The graph shows the percentage of attached cells compared with the control group. <b>C</b> Western blot analysis of the relative protein levels of Src and FAK in BRMS1 overexpression and control group for both U251 and U87 cell lines. <b>D</b> Quantitative analysis of relative protein level of Src and FAK in glioma U251 and U87 cells. All experiments were carried out in triplicate. Data are shown as mean ± SD. *<i>P</i><0.05, **<i>P</i><0.01.</p

Representative images show BRMS1 immunohistochemical staining.

<p><b>A</b> Positive BRMS1 staining in normal brain tissue (NB); <b>B</b> Positive BRMS1 staining in adjacent normal brain tissue (AB); <b>C</b> Negative BRMS1 staining in benign tumor (BT); <b>D</b> Negative BRMS1 staining in malignant tumor (MT); <b>E</b> A significant difference in BRMS1 staining was observed between normal brain tissue and glioma tissue (GT) (<i>P</i><0.01, χ² test) and between tumor adjacent normal brain tissue and glioma tumor (<i>P</i><0.01, χ² test). <b>F</b> Whole-cell protein extracts were further prepared from four paired tumor adjacent normal glioma tissues and glioma tissues. The BRMS1 protein level was determined by Western blot analysis. <b>G</b> Western blot analysis of BRMS1 expression in normal human astrocytes NHA and glioma cell lines, including SHG44, C6, U251, T98G, U87. <b>H</b> BRMS1 staining was dramatically decreased in malignant tumor compared with benign tumor (<i>P</i><0.05). All experiments were carried out in triplicate. Data are shown as mean ± SD. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001. Original magnification (<b>A–D</b>) ×40.</p

Overexpression of BRMS1 inhibits glioma cell migration.

<p><b>A, B</b> Wound-healing assay was done on monolayers of U251and U87 glioma cells after 24 hour of transfection. The photographs were taken at 0 and 24 hours after wounds were made. <b>C, D</b> Cell migration assay was performed after overexpression of BRMS1 in glioma U251 and U87 cells. The graph shows the percentage of cells migrated in 5 fields of view compared with the control group. All experiments were carried out in triplicate. Data are shown as mean ± SD. **<i>P</i><0.01; ***<i>P</i><0.001.</p

Overexpression of BRMS1 suppresses cell invasion but not cell proliferation in glioma cells.

<p><b>A</b> Twenty-four hours after transfection, the expression of BRMS1 in U251 and U87 glioma cells was evaluated by western blot. <b>B, C</b> CCK-8 cell proliferation assay was performed after BRMS1 overexpression in U251 and U87cells. <b>D, E</b> Matrigel cell invasion assay was performed after the overexpression of BRMS1 in U251 and U87 cells. The graph shows the percentage of cells invaded in 5 fields of view compared with the control group. <b>F</b> BRMS1 inhibits MMP-2 activity in U251 and U87 cells by zymography. <b>G</b> Western blot analysis of the relative protein levels of MMP-2, uPA and NF-κB in BRMS1 overexpression and control group of U251 and U87 cells. <b>H</b> Quantitative analysis of relative protein level of MMP-2, uPA, p27 and NF-κB in glioma U251 and U87 cells. All experiments were carried out in triplicate. Data are shown as mean ± SD. *<i>P</i><0.05, **<i>P</i><0.01.</p