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Fig. 3 | BMC Cancer

Fig. 3

From: Planar cell polarity gene expression correlates with tumor cell viability and prognostic outcome in neuroblastoma

Fig. 3

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 < 0.05, **P < 0.01

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