Project description:Four cervical carcinoma cell lines (C4I, CaSki, HeLa, SiHa) were treated with epigenetic modifying drugs 5-aza-2’-deoxycytidine and trichostatin A. Such treatment leads to reactivation of genes that are epigenetically silenced in cancer by aberrant methylation; reactivated genes were then identified by microarray profiling. Subsets consist of 2 biological repeats and 2 dye-flip experiments for each repeat for CaSki cell line, and 3 repeats, each with 2 dye-flip experiments for C4I, HeLa, Caski cell lines. The identification of reactivated genes, their selection for validation experiments, and final identification of 6 genes that are significantly more often hypermethylated in cervical carcinoma clinical specimens is described in (manuscript submitted). Keywords: other
Project description:We have sequenced miRNA libraries from human embryonic, neural and foetal mesenchymal stem cells. We report that the majority of miRNA genes encode mature isomers that vary in size by one or more bases at the 3’ and/or 5’ end of the miRNA. Northern blotting for individual miRNAs showed that the proportions of isomiRs expressed by a single miRNA gene often differ between cell and tissue types. IsomiRs were readily co-immunoprecipitated with Argonaute proteins in vivo and were active in luciferase assays, indicating that they are functional. Bioinformatics analysis predicts substantial differences in targeting between miRNAs with minor 5’ differences and in support of this we report that a 5’ isomiR-9-1 gained the ability to inhibit the expression of DNMT3B and NCAM2 but lost the ability to inhibit CDH1 in vitro. This result was confirmed by the use of isomiR-specific sponges. Our analysis of the miRGator database indicates that a small percentage of human miRNA genes express isomiRs as the dominant transcript in certain cell types and analysis of miRBase shows that 5’ isomiRs have replaced canonical miRNAs many times during evolution. This strongly indicates that isomiRs are of functional importance and have contributed to the evolution of miRNA genes
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
