Figure 6.

Statins decreased β-catenin target genes. (A) Simvastatin and lovastatin attenuated AXIN-2 mRNA, as a β-catenin target gene, in lung cancer and melanoma cells. Lung cancer (NCI-H460 and Calu-1) and melanoma (A2058) cells were incubated with simvastatin (5 µM) or lovastatin (5 µM) for 6 h, as indicated. AXIN-2 was measured by using qRT-PCR. Experiments were performed in triplicates. All plots indicate the mean ± SD. ** p < 0.01 and *** p < 0.001 by one-way ANOVA, followed by an appropriate post hoc test for the comparison between two experimental groups. (B) Chromatin immunoprecipitation (ChIP) for β-catenin followed by ChIP-qPCR in A2058 and Calu-1 cells confirmed a chromatin enrichment of β-catenin at the CD274 promoter loci (600 bp upstream from the transcription start site). Cells were incubated for 6 h in the absence or presence of 25-HC (20 µM) and then cells were further incubated with 5 µM of simvastatin for 8 h. All plots indicate the mean ± SD. Experiments were performed in triplicates. * p < 0.05 and ** p < 0.01 by one-way ANOVA, followed by an appropriate post hoc test for the comparison between two experimental groups. (C) β-catenin target genes, such as CCND1, BIRC5, ENC1, MMP7, AXIN-2, BCL2, and TP53, mRNA expression levels were measured in pravastatin-treated Calu-1 cells using a publicly available microarray dataset (GSE47458). All plots indicate the mean ± SD. ** p < 0.01 by Student’s t-test for two experimental comparisons. (D,E) CCND1, MMP2, BIRC5, ID2, ENC1, and MMP7 mRNA levels were measured in simvastatin, lovastatin, or fluvastatin-treated (D) PANC1 and (E) MiaPaca2 pancreatic cancer cells using a publicly available microarray dataset (GSE149566). All plots indicate the mean ± SE. * p < 0.05 and ** p < 0.01 by one-way ANOVA, followed by an appropriate post hoc test for the comparison between two experimental groups.