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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Differential gene expression profile of y79 retinoblastoma cells in response to Anti - HMGA2 siRNA treatment

ABSTRACT: The differential gene expression study was conducted in the Anti - HMGA2 siRNA treated Retinoblastoma cell (Y79). The transient transfection in Y79 cells were performed using Anti-HMGA2 siRNA . The concentration and the time of incubation for the transfection was optimised to increase the silencing efficiency and reduce the off -target effects. After 48 hours of transfection the cells were collected and the whole genome cDNA microarray was performed to interpret the differential gene expression regulated by HMGA2 in Y79 cells. The results of the microarray data has been validated. One-color experiment,Organism: Human , Custom Whole Genome Human 8x60k designed by Genotypic Technology Pvt. Ltd. (Agilent-027114), Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)

ORGANISM(S): Homo sapiens

SUBMITTER: Genotypic technology

PROVIDER: E-GEOD-31687 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Differential gene expression profile of y79 retinoblastoma cells in response to Anti - HMGA2 siRNA treatment

Project description:The differential gene expression study was conducted in the Anti - HMGA2 siRNA treated Retinoblastoma cell (Y79). The transient transfection in Y79 cells were performed using Anti-HMGA2 siRNA . The concentration and the time of incubation for the transfection was optimised to increase the silencing efficiency and reduce the off -target effects. After 48 hours of transfection the cells were collected and the whole genome cDNA microarray was performed to interpret the differential gene expression regulated by HMGA2 in Y79 cells. The results of the microarray data has been validated.

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Differential expression of miRNAs in Retinoblastoma cells (Y79) in response to anti-HMGA2 siRNA treatment

Project description:The main objective of the study is to identify the de-regulated miRNAs in association with HMGA2 (oncogene) in the Retinoblastoma tumorus. The present experiment was carried out using Y79 (Retinoblastoma cell line), as the invitro model of Retinoblastoma tumor. The HMGA2 transcripts were silenced using siRNA and the post-silenced RB cells (at the end of 48 hours) were subjected to miRNA profiling using agilent platform.The key miRNAs de-regulated in this experiment has been validated using qRT-PCR and further their role has been evaluated by addition of specific antagomirs in RB cell lines (Y79 and Weri Rb 1). Agilent one-color experiment,Organism: Homo sapiens ,Agilent Human miRNA 8x15k Arrays AMADID: 021827 [Agilent miRNA labeling reagent and Hybridization Kit Cat # 5190-0408] RB cells (Y79) treated with anti-HMGA2 siRNA or with control

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Project description:We have reported that the nuclear protein high mobility group A2 (HMGA2), is required for the induction of EMT by TGF-beta (Thuault et al. J. Cell Biol. 2006). HMGA2 regulates a cohort of transcriptional regulators of the EMT process, such as Snail1 and Twist1 (Thuault et al. J. Biol. Chem. 2008, E-Jean Tan et al. J. Biol. Chem. 2012). Since HMGA2 is a chromatin-associated protein, we expect that it should regulate many genes in addition to the EMT-related factors. The aim of our study is to decipher novel molecular mechanisms that explain how EMT links to the process of tumor-initiating cell self-renewal via the chromatin factor HMGA2. In this experiment, we stably transfected human breast cancer MDA-MB-231 cells with shRNA against HMGA2 and investigated effects of the knock-down on global gene expression using Affymetrix arrays, in order to identify novel downstream players involved in this network. In parallel experiments we also analyzed the phenotypic effects of the HMGA2 knock-down on MDA-MB-231 cell proliferation, survival and inavasiveness.

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