Project description:To determine the molecular basis for CD99 function in osteosarcoma, gene expression profile of U-2/CD99 wild-type(wt) expressing cell lines were compared with U-2 OS parental cells.
Project description:Disassembly of the nuclear envelope is an essential feature of mammalian cell division controlled by the phosphoryaltion of lamins via cyclin dependent kinases. This process is affected in cells expressing progerin, a lamin A allele found in patients with Hutchinson-Gilford Progeria syndrome. Progerin can inhibit cell proliferation of both normal and tumor cells and this property is largely magnified if its phosphorylation at serine 22 is inhibited by a genetic mutation to generate S22A-progerin. Surprisingly, S22A-progerin acquires the ability to trigger cellular senescence in tumor cells with mutations in the p53 and RB tumor suppression pathways suggesting a novel pathway to control the growth of malignant tumors. We used microarrays to characterize the gene expression changes induced by the S22A-progerin in comparison with the wt-progerin in U-2 OS cell line. U-2 OS cells were infected with a retroviral vector that express S22A-progerin or wt-progerin. After 4 days of infection, RNA was extracted from three independents replicas of each condition. Total RNA was send to Genome Quebec service for hybridization with Affymetrix microarrays.
Project description:expression analysis from a genetically engineered mouse model of osteosarcoma; determine the expression profile of mouse osteosarcoma Experiment Overall Design: 3 control in vitro differentiated WT primary osteoblasts; 15 primary osteosarcoma; 4 OS cell lines; 4 secondary tumours
Project description:Disassembly of the nuclear envelope is an essential feature of mammalian cell division controlled by the phosphoryaltion of lamins via cyclin dependent kinases. This process is affected in cells expressing progerin, a lamin A allele found in patients with Hutchinson-Gilford Progeria syndrome. Progerin can inhibit cell proliferation of both normal and tumor cells and this property is largely magnified if its phosphorylation at serine 22 is inhibited by a genetic mutation to generate S22A-progerin. Surprisingly, S22A-progerin acquires the ability to trigger cellular senescence in tumor cells with mutations in the p53 and RB tumor suppression pathways suggesting a novel pathway to control the growth of malignant tumors. We used microarrays to characterize the gene expression changes induced by the S22A-progerin in comparison with the wt-progerin in U-2 OS cell line.
Project description:Osteosarcoma (OS) is a common primary bone malignancy that is characterized by high degree of aneuploidy, gene amplification, and multiple unbalanced chromosomal rearrangements. The human osteosarcoma U-2 OS and Sa OS cells lines have been generated more than three decades ago and are used in a wide spectrum of biomedical research. Nevertheless, and despite scattered information about their genetic context, no comprehensive comparative study of their transcriptome profile has been reported to date. The aim of this study was to elucidate common molecular characteristics of the two cell lines as well as differences in their expression profile. Thus the genome wide gene expression profile of the Sa OS cells was compared with reference RNA of U-2 OS cells. These results may provide the basis for future studies and illuminate the molecular differences of the two widely used cell lines.
Project description:Here we made use of a human osteosarcoma U-2 OS modified cell line (U-2 OS DIvA- Flag-GFP-SMC1), expressing a hydroxytamoxifen controlled restriction enzyme (AsiSI-ER which allows the induction of multiple annotated DSBs) and Flag GFP-SMC. We performed tandem purification before and after DSB induction followed by mass spectrometry to identify cohesin interacting partners.
