Journal: BMC Cancer
Article Title: Planar cell polarity gene expression correlates with tumor cell viability and prognostic outcome in neuroblastoma
Figure Lengend Snippet: Inhibition of Wnt/PCP downstream effector ROCK increases Prickle1 expression and represses active β-catenin. a , b mRNA expression of Prickle1 and Vangl2 after treatment with ROCK inhibitor HA1077 or Y27632 for 72 h; results showed a consistent increase in Prickle1 expression (one-way ANOVA with Bonferroni post-test, SK-N-AS Prickle1: P = 0.0017, control vs HA1077 50 μM P = 0.0023, control vs HA1077 50 μM P = 0.0021, Vangl2: P = 0.0061, control vs HA1077 50 μM P = 0.017 and SH-SY5Y Prickle1 P = 0.019, control vs HA1077 50 μM P = 0.035, control vs HA1077 50 μM P = 0.020 and t -test, SK-N-BE (2) Vangl2 control vs Y27632 80 μM P = 0.0015). Expression of mRNA (relative to the vehicle treated control normalized to the mean expression of the housekeeping genes) was determined by real-time RT-PCR, means with S.D. of triplicates are displayed. c Transcriptional activity of β-catenin after HA1077 exposure; cells were transfected with a TCF/LEF luciferase reporter construct and treated with HA1077 (25 or 50 μM). TOPFlash-dependent activity was significantly reduced as compared with the control (one-way ANOVA with Bonferroni post-test: SK-N-AS P = 0.0144, control vs HA1077 50 μM P = 0.029 and SK-N-BE (2) P = 0.0023, control vs HA1077 50 μM P = 0.012, control vs HA1077 50 μM P = 0.0017). Data represent the mean and SD of three determinations and the experiment was repeated twice. d Protein expression of active β-catenin following HA1077 exposure (96 h, HA1077 25 or 50 μM), determined by western blotting. e mRNA expression of Prickle1 and Vangl2 after knockdown of β-catenin in neuroblastoma cells SK-N-AS and SK-N-BE(2), no significant changes were observed. * P
Article Snippet: The mRNA expression of Prickle1 was not affected after knockdown or overexpression of Vangl2 and similarly, the mRNA expression of Vangl2 was not affected after knockdown or overexpression of Prickle1 in SK-N-AS or SK-N-BE (2) cells (Fig. ).
Techniques: Inhibition, Expressing, Quantitative RT-PCR, Activity Assay, Transfection, Luciferase, Construct, Western Blot