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| Status |
Public on Aug 20, 2013 |
| Title |
Knockdown of human TCF4 affects multiple signaling pathways involved in cell survival, epithelial-to-mesenchymal transition and neuronal differentiation |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
TCF4 is an important neurodevelopmental transcription factor associated with schizophrenia and Pitt-Hopkins syndrome. In this study, we used genome-wide expression profiling to determine the effects of acute TCF4 knockdown on gene expression in SH-SY5Y neuroblastoma cells. Pathway and enrichment analysis on the differentially expressed genes in TCF4-knockdown cells identified an over-representation of genes involved in TGF-β signaling, epithelial-to-mesenchymal transition (EMT) and apoptosis. Among the most significantly differentially expressed genes were the EMT regulators SNAI2 and DEC1 and the proneural genes NEUROG2 and ASCL1. Altered expression of several mental retardation genes such as UBE3A (AS), ZEB2 (MWS) and MEF2C was also found in TCF4-depleted cells. These data suggest that TCF4 regulates a number of convergent signaling pathways involved in cell differentiation and survival in addition to a subset of clinically important mental retardation genes.
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| Overall design |
To identify TCF4-mediated gene expression changes in SH-SY5Y cells, experiments were conducted with two control (mock and GAPDH KD) and two TCF4 knockdown (KD1 and KD2) groups. Two non-overlapping siRNAs against TCF4 were designed and transfected over a 72h period to induce global knockdown of TCF4 transcripts. In parallel, cells were mock transfected (mock; transfection reagent only) and transfected with an unrelated siRNA against GAPDH (GAPDH KD) in order to control for background gene expression changes due to transfection, presence of dsRNA and activation of the cellular silencing machinery. After transfection, RNA was extracted from all cells, converted to cDNA and labelled for microarray hybridization. Each of the four treatment groups (mock, GAPDH KD, KD1 and KD2) contained three biological replicates which were processed at the same time using the Toray microarray platform.
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| Contributor(s) |
Forrest MP, Waite AJ, Martin-Rendon E, Blake DJ |
| Citation(s) |
24058414 |
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| Submission date |
Jun 27, 2013 |
| Last update date |
Sep 24, 2013 |
| Contact name |
Derek Blake |
| E-mail(s) |
[email protected]
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| Organization name |
Cardiff University
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| Department |
MRC Centre for Neuropsychiatric Genetics and Genomics
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| Lab |
Blake Lab
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| Street address |
Hadyn Ellis Building, Maindy Road
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| City |
Cardiff |
| ZIP/Postal code |
CF24 4HQ |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL13915 |
3D-Gene Human Oligo chip 25k V2.1 |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA209851 |
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