Figure 5
From: Regeneration linked miRNA modify tumor phenotype and can enforce multi-lineage growth arrest in vivo
DNMT1 expression in HepG2 and RKO cells transfected with miRNA constructs, methylation and gene expression arrays in miRNA-152i generated xenograft tumors and expression of genes associated with cell proliferation and cancer in HepG2 and RKO cells transfected with miRNA constructs. (a) qPCR analysis of DNMT1 expression. (i) HepG2 and (ii) RKO cells transfected with miRNA constructs. Kidney RNA used as a positive control. All expression levels were normalized to scrambled control vector. qPCR data in all cases are representative of three different experiments. Error bars indicate SD. *p < 0.05, **p < 0.01, ***p < 0.001. (b) Analysis of the total CpG methylation in cells transduced with scrambled control vector and miRNA-152i. The datasets are equivalent, overlapping and not skewed (3% difference); (i): box and whisker plot, (ii) principle component analysis (PCA) of the degree of methylation at each CpG in the two datasets shows samples group by condition. (c) Gene expression analysis using Affymetrix gene2 array: RNA extracted from xenograft tumors transduced with scrambled or miRNA-152i. (i) PCA shows complete segregation of 2 groups indicating differentially expressed genes within each group, (ii)The dendrogram shown above heatmap demonstrates similarities and differences between samples by miRNA expression (Affymetrix GeneChipm Command Console (AGCC) 4.0, http://www.affymetrix.com/support/technical/byproduct.affx?product=commandconsole). (d) qPCR analysis of SNRPN, WNK3, and FAM3B genes. (i) HepG2 and (ii) RKO) cells transfected with scrambled, miRNA-150, miRNA-152i and miRNA-150/-152i. All expression levels were normalized to scrambled control vector. qPCR data in all cases are representative of three different experiments. Error bars indicate SD. *p < 0.05, **p < 0.01, ***p < 0.001.
