Project description:The transcriptome-wide response of hypoxic breast cancer cells upon MBNL2 knockdown was analyzed. Differential gene expression and alternative splicing were investigated.
Project description:While analyzing mRNA expression profiles of clear cell renal cell carcinoma (ccRCC) tumors, we found that the mRNAs that are bound at their 3' UTRs by muscleblind-like splicing regulator 2 (Mbnl2) and epithelial splicing regulatory protein 2 (ESRP2) are up-regulated in tumor compared to patient-matched normal tissues. Given that MBNL2 increases the stability of its targets and ESRP2 destabilizes its targets, we predicted that, in ccRCC tumors, MBNL2 activity increases, while ESRP2 activity decreases. To investigate the effect of each of these two RNA-binding proteins (RBPs) on the transcriptome of the cell, we used shRNA to knockdown MBNL2 in two different cell line models of ccRCC (786-O and A-498), and also to knockdown ESRP2 in normal primary renal proximal tubule epithelial cells (PRPTEC). RNA-seq revealed that MBNL2 knockdown partially reverses the ccRCC-associated transcriptome in ccRCC cell lines, whereas ESRP2 knockdown shifts the transcriptome of PRPTEC toward that of ccRCC.
Project description:Experiments were performed assessing whether targeting the pH regulatory proteins (CAIX, NHE1 and V-ATPase) that permit cancer cells to adapt to hypoxic conditions could produce an effective therapeutic response in breast cancer, using both 2D and 3D culture models. CAIX inhibitors were shown to combine effectively with irradiation in clonogenic assays. Proteomic-mass-spectrometric analysis was carried out to analyze the possible mechanisms through which the CAIX inhibitor used was combining with irradiation.
Project description:G9a is able to silence gene expression in hypoxic condition by increasing histone H3K9me2. We have identified a set of genes that are negatively regulated by G9a in hypoxia-dependent manner. In this dataset, we include the expression data obtained from MCF7 breast epithelial cells that have been transfected with control (WT) or G9a shRNAs (KD) and exposed to either normoxia or hypoxia. These data are used to obtain 829 genes that are differentially expressed in response to hypoxia, and 205 genes that are sentisive to G9a level. 4 samples were analysed. We generated comparisons between WT and KD in normoxic as well as hypoxic condition. Genes differentially expressed in hypoxic condition were selected followed by selection of genes that lose this differential expression upon G9a knockdown.
Project description:The study aims to elucidate the effect of histone methyltransferase SMYD3 on gene expression in MCF-7 breast cancer cell line. Knockdown luciferase control v.s. knockdown SMYD3 in MCF-7 breast cancer cell line were conducted. Results identify a large proportion of cell cycle-related genes regulated by SMYD3.