5 research outputs found
Analysis of 45S rDNA promoter methylation and expression of rRNA transcripts in breast cancer
Cataloged from PDF version of article.Ribosome biogenesis has a central role in cell growth and proliferation that is usually
disrupted in tumor cells by the inactivation of tumor suppressor genes and activation
of oncogenes. Ribosomal RNA (rRNA) gene expression is one of the most important
factors regulating ribosome production, which is controlled by CG rich 45S
ribosomal DNA (rDNA) promoter. The effect of DNA methylation at 45S rDNA
promoter on rRNA gene expression is a subject of controversy in the literature. In
this thesis, a 434 bp region (-380 bp to +54 bp) spanning both upstream control
element (UCE) and core promoter located in 45S rDNA promoter containing 54
CpGs was analysed in breast cancer. We also analysed the related rRNA expression
levels in the same samples in order to clarify the role of 45S rDNA promoter
methylation on rRNA gene expression.
45S rDNA promoter region was highly methylated (74%-96%) in all cell lines
including non-tumorigenic breast cell line (MCF10A). Even though 45S rDNA
promoter region of breast cancer cell lines are extensively methylated, rRNAs (18S,
28S, 5.8S and 45S ETS) were expressed independent of the heavy methylation.
Expression levels of rRNAs are assessed either using housekeeping genes (ACTB,
TBP, ACTB&TBP) or geometric mean of rRNAs (GM-rRNAs). We propose GMrRNA
normalization as a new method to identify relative expression differences
between rRNA transcripts.
Epigenetic drugs 5-Aza-2’-deoxycytidine (5-AZA) and Trichostatin A (TSA) were
used to determine the effect of DNA methylation and histone acetylation on rRNA
expression. Demethylation with 5-AZA resulted in an unexpected decrease in the
expression of all rRNA. TSA treatment did not lead to any significant expression
difference in cell lines.
To better evaluate the effect of DNA methylation on the expression of rRNA
transcripts we analysed the methylation status of 19 breast tumor and matched
normal frozen tissue samples. The results showed that majority of the tumors (13/19)
have significantly higher methylation levels than their normal pairs. Using the GMrRNA
as reference helped us to determine significant differences in the proportionate
expression of rRNAs in these tissue samples. The 5.8S rRNA ratio was significantly
lower whereas the 18S rRNA ratio was significantly higher in breast tumor samples.
Furthermore, the 45S rDNA promoter methylation levels in normal breast tissue
samples were negatively correlated with the18S rRNA ratio but this correlation was
disrupted in breast tumors. Similarly, rRNA transcript levels were significantly
correlated with each other in normal samples, were lost in tumor samples. It is clear
that, there is a dysregulation both in rDNA methylation levels and spliced rRNA
transcripts specific to breast tumor samples, which was not observed in normal breast
tissues. rRNA gene expression is controlled by mechanisms other than promoter
DNA methylation. Tumorigenesis may cause disruption of many control mechanisms
that are required for proper rRNA expression, splicing and maturation, resulting in a
dysregulation of the correlation between spliced rRNA expression levels, which
should be investigated further.Karahan, GurbetPh.D
Transgelin gene is frequently downregulated by promoter DNA hypermethylation in breast cancer
Tables not included in the main manuscript have been listed. Table S1. Number of probe sets affected by AZA treatment; Table S2. Comparison of significantly altered probe sets with the independent study GSE20713 Dataset; Table S3. Cancer vs. normal analysis of TAGLN mRNA in Oncomine database. (PDF 18Ă‚Â kb
Journée d'étude : L’usager, l’assistance et la question de la contrepartie
Journée en hommage à Robert Castel organisée par le LIRTES (UPEC) Le mardi 20 mai 2014 à Créteil Avec la participation de Colette Bec (« Usages politiques de l’assistance en démocratie ») et Christophe Trombert (« L’assistance aux pauvres valides : évolutions des contreparties »), membres du laboratoire LISE Bien que l’usager soit une catégorie relativement ancienne servant à désigner le rapport entre les personnes qui utilisent un service (logement, école, transport…) et les pouvoirs publics..
Additional file 3: of Transgelin gene is frequently downregulated by promoter DNA hypermethylation in breast cancer
Final gene lists altered upon AZA treatments only in breast cancer cell lines. Probe IDs, gene names, accession numbers and fold change ratios of the genes altered upon AZA treatment, in each breast cancer cell line at 1.5 fold and P < 0.05, and filtered as shown in Fig. 1A have been listed. (XLSX 387 kb