Figure 1From: Epigenetic silencing of RASSF1A deregulates cytoskeleton and promotes malignant behavior of adrenocortical carcinomaRASSF1A expression and regulation in adrenal tumor. (A) Averages of percentage methylated (FM) and unmethylated (FUM) CpGs in CpG island A of RASSF1A promoters in Normal adrenal cortex (n = 6), ACA (n = 8), and ACC (n = 7) samples are shown. FM includes both Hypermethylated (FHM) and intermediate methylated (FIM) fractions. (B) Methylation profiles of individual fresh-frozen normal adrenal cortex (N1 – N6), 8 ACAs (A1 – A8) and 7 ACC samples (C1 – C7) as determined by Epitect methyl II PCR assay. (C) Expression of RASSF1A mRNA determined by real-time qPCR in 7 ACC samples (C1 – C7) compared to the average expression in 6 normal samples (N) normalized to a value of 1.0. RASSF1A expressions in individual samples were also normalized to the average mRNA expression of house-keeping genes beta-actin (Actb) and TATA-binding protein (TBP). C-Av represents the average expression of all ACC samples. Data shown is from one of triplicate experiments that yielded similar results (mean ± SD). Independent sample t-test used to derive the p value (p = <0.01). (D) RASSF1A protein expression in normal (a &b) and ACC (c &d) FFPE tissue specimens demonstrated by immunohistochemistry through DAB staining (brown indicates RASSF1A protein expression) followed by nuclear counterstaining by hematoxylin (blue).Back to article page