Aberrant Hypermethylation-Mediated Suppression of PYCARD Is Extremely Frequent in Prostate Cancer with Gleason Score ≥ 7

Figure 1

Effects of NFκB (AD)-MBD induction on cell proliferation, apoptosis, and PYCARD upregulation. (a) Results of cell proliferation assays for LNCaP-derived TR9, TR9-MeTA14, TR15, and TR15-MeTA5. and denote and , respectively. The photos below show the light microscopic appearance (40x magnification). Results of flow cytometry (b) and TUNEL ((c), 200x magnification) assays. (d) Results of RT-PCR analysis of PYCARD and TNFRSF25 genes. (e) Methylation statuses obtained from bisulfite sequencing of PYCARD and TNFRSF25 promoter regions are schematically shown. Closed and open circles indicate the methylated and unmethylated CpG sites, respectively. Red arrows indicate the transcriptional start site (TSS) at position +1. Incidental genetic alterations were found in LNCaP, in TNFRSF25, a G to A transition at position -64 (indicated by open triangles) and two deletions (indicated by closed triangles), one between +10 and +13 (4 bp) and the other between +46 and +53 (8 bp). These alterations contain one CpG site.

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