Journal: PLoS ONE

Article Title: A Rac1/Cdc42 GTPase-Specific Small Molecule Inhibitor Suppresses Growth of Primary Human Prostate Cancer Xenografts and Prolongs Survival in Mice

doi: 10.1371/journal.pone.0074924

Figure Lengend Snippet: Analysis of PAK, AKT and BAD-phosphorylation in EGF-stimulated 22Rv1 prostate cancer cells following AZA1 treatment. Representative Western blot images and quantification of immunoblots stained with phospho-PAK1/2 (pPAK), phospho-AKT (pAKT) and phospho-BAD (pBAD) antibodies before and after treatment with 2, 5 and 10 µM AZA1 for 24 h. Rac1/Cdc42 blockade reduces phosphorylation of PAK1, AKT and BAD in 22Rv1 prostate cancer cells compared to controls (means of 3 independent experiments). *, significantly different from unstimulated and untreated controls; + , significantly different from EGF-stimulated control; ‡ , significantly different from unstimulated, untreated control and EGF-stimulated control. SP, specific protein; LC, loading control.

Article Snippet: The blots were probed with antibodies against phospho-PAK1 (pS144)/PAK2 (pS141) (Cell Signaling Technology, Danvers, MA), phospho-AKT (anti-phospho-AKT pT308 from BD Biosciences) and phospho-BAD (anti-phospho-BAD Ser112 from Cell Signaling Technology) before incubation with horseradish peroxidase–conjugated secondary antibodies (Amersham Pharmacia Biotech).

Techniques: Phospho-proteomics, Western Blot, Staining, Control

Journal: PLoS ONE

Article Title: A Rac1/Cdc42 GTPase-Specific Small Molecule Inhibitor Suppresses Growth of Primary Human Prostate Cancer Xenografts and Prolongs Survival in Mice

doi: 10.1371/journal.pone.0074924

Figure Lengend Snippet: AZA1 inhibits activation of Rac1 and Cdc42 GTPases, shifting the balance towards cell growth inhibition and apoptosis. These effects are exerted by different mechanisms. AZA1 inhibits the PAK pathway via the cell cycle regulator Cyclin D1 and suppresses PAK and AKT activation leading to reduced BAD phosphorylation to induce pro-apoptotic activity. In addition, suppression of Rac1 and Cdc42 suppresses cancer cell migration and invasion by affecting actin polymerization and subsequent inhibition of lamellipodia and filopodia formation.

Article Snippet: The blots were probed with antibodies against phospho-PAK1 (pS144)/PAK2 (pS141) (Cell Signaling Technology, Danvers, MA), phospho-AKT (anti-phospho-AKT pT308 from BD Biosciences) and phospho-BAD (anti-phospho-BAD Ser112 from Cell Signaling Technology) before incubation with horseradish peroxidase–conjugated secondary antibodies (Amersham Pharmacia Biotech).

Techniques: Activation Assay, Inhibition, Phospho-proteomics, Activity Assay, Migration