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RNA-seq with knockdown ZBTB7A and NTC control

ABSTRACT: RNA-seq with knockdown ZBTB7A and NTC control

PROVIDER: PRJNA507693 | ENA |

REPOSITORIES: ENA

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RNA-seq with knockdown ZBTB7A and NTC control

Project description:Zinc finger and BTB domain containing transcription repressor ZBTB7A has been recently reported as a tumor suppressor who plays important functions to prevent the progression of prostate cancer. However, the chromatin activity of ZBTB7A in prostate cancer cells remain unclear. In order to identify the cistrome and transcriptome of ZBTB7A, we performed ZBTB7A ChIP-seq in VCaP cells and RNA-seq in VCaP cells transfected with siRNA targeting ZBTB7A or non-targeting control, respectively (cells were grown in full serum). By using combined ChIP-seq and RNA-seq analyses in VCaP cells, we have precisely mapped the ZBTB7A binding sites and identified a subset of genes that are directly repressed by ZBTB7A. To study the chromatin interaction of ZBTB7A with androgen receptor (AR), we also performed ChIP-seq of ZBTB7A in VCaP cells stimulated with 10 nM DHT for 4 hours versus vehicle control (cells were hormone-depleted prior to DHT stimulation). Our results show that ZBTB7A binding is increased by androgen at AR and ZBTB7A overlapping sites.

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ZBTB7A ChIP-seq under different DHT level

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RNA Sequencing Quantitative Analysis of control and knockdown ZBTB7A.

Project description:The aim of the study is to compare the RNA-sequencing data to protein array and qRT-PCR and to clarify the effect of ZBTB7A on apoptosis.

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Project description:Tumor budding (TB), clusters of less than 5 cells dissociated from the tumor mass, is reported to predict nodal involvement and recurrence in multiple human cancers including gastric carcinoma (GC). However, it is not clear how TB forms. In this study, we determined the expression pattern of GAGE12I, CTNND1, Kif26B, CREB1 and ZBTB7A in human GC samples by IHC attempting to find any TB-related markers. The results showed that TB could predict lymph node metastasis and is negatively associated with the overall survival of GC patients. The expression of ZBTB7A in the tumor margin (invasive front), rather than the other four markers, was much higher than that inside the tumor and was positively correlated with TB score. Moreover, patients with higher marginal ZBTB7A expression had poorer prognosis. ZBTB7A could enhance the migration and invasion capabilities of GC cells in vitro. Mechanically, ZBTB7A might activate EGFR-MAPK-ERK and PI3K-AKT-mTOR pathway, which resulted in epithelial-mesenchymal transition (EMT) to induce the formation of TB. Thus, we concluded that higher ZBTB7A expression in the tumor margin may contribute to the dissociation of tumor cells from the tumor mass to form TB by initiating EMT via EGFR-MEK-ERK and PI3K-AKT-mTOR pathway.

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Project description:Mass spectrometry of wild type or mutant (RNA-binding) ORC1. Experiments were done in cellstransfected with a control ASO or an anti-GAA ASO, to knockdown RNA interactors of ORC1.

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