Journal: Molecular Cancer

Article Title: Arecoline induces HA22T/VGH hepatoma cells to undergo anoikis - involvement of STAT3 and RhoA activation

doi: 10.1186/1476-4598-9-126

Figure Lengend Snippet: Arecoline alters the expression of apoptosis-related proteins and caspase activity in HA22T/VGH cells . (A) HA22T/VGH cells were treated with 0, 30, or 100 μg/ml of arecoline for 24 h, then the cells were harvested and proteins extracted and used for Western blotting for Bcl-2, Bcl-X L , Bax, cytochrome c, or procaspase-9. β-actin was used as the internal control. The values shown are the quantitative density analysis expressed as the relative density compared to that in untreated cells (control), taken as 100%. The results are expressed as the mean ± S.D. for three separate experiments. (B) Caspase-3 activity was detected using RPE-conjugated anti-active caspase-3 antibody by flow cytometric analysis. The values shown are the percentage of cells with active caspase-3 and are the mean ± S.D. of three independent experiments. The red filled area is the untreated control and the black lines the treated groups. *: p < 0.05 as compared to the untreated control.

Article Snippet: R-phycoerythrin (RPE)-conjugated rabbit anti-active caspase-3 polyclonal antibodies, RPE-conjugated mouse anti-human β1-integrin monoclonal antibody and the RPE-conjugated mouse IgG isotype control were purchased from BD Pharmingen Inc. (San Diego, CA, USA).

Techniques: Expressing, Activity Assay, Western Blot

Journal: Molecular Cancer

Article Title: Arecoline induces HA22T/VGH hepatoma cells to undergo anoikis - involvement of STAT3 and RhoA activation

doi: 10.1186/1476-4598-9-126

Figure Lengend Snippet: Schematic representation of the arecoline-stimulated signaling pathways for detachment and apoptosis of HA22T/VGH cells . Arecoline treatment decreases IL-6 levels, but does not change gp130 of IL-6 receptor. In addition, phosphorylation/activation of STAT3, which provides protection against anoikis, is inhibited and levels of its downstream signals IL-6, Bcl-2, and Bcl-X L are decreased, while Bax levels, mitochondrial cytochrome c release, caspase-9 levels, and caspase-3 activity are increased. Phosphorylation/activation of p190RhoGAP, a RhoA inhibitor, and its upstream regulator, SHP2, are inhibited, while the activation/cleavage of Rock-1, a RhoA downstream kinase, and actin stress fiber formation are increased, contributing to anoikis.

Article Snippet: R-phycoerythrin (RPE)-conjugated rabbit anti-active caspase-3 polyclonal antibodies, RPE-conjugated mouse anti-human β1-integrin monoclonal antibody and the RPE-conjugated mouse IgG isotype control were purchased from BD Pharmingen Inc. (San Diego, CA, USA).

Techniques: Activation Assay, Activity Assay