Regualtion of 5hMC during Twist-indced EMT
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ABSTRACT:
PROVIDER: PRJNA943282 | ENA |
REPOSITORIES: ENA
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Project description:EMT is associated with profound epigenetic regualtion. 5hMC is the first product of TET-dependent DNA methylation product. chromatin immunoprecipitation-sequencing (ChIP-seq) provide a tool to define the changes of 5hmC at the genome level. Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for 5hMC in HMLE cells.
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Project description:The power of our genome-wide survey lies in that we were able to query chromatin accessibility changes concurrently with those in DNA methylation. We identified both opening and closing events, predominantly independent of DNA methylation, in both systems. Our results have powerful implications for future basic and clinical research on this developmental process and its resurfacing during cancer cell metastasis. To address epigenetic changes that mediate EMT, we used two models in the human system: EMT induced by the transcriptional repressor Twist as well as EMT induced by TGF-beta. We performed AcceSssIble on these cells at several time points through the course of the transition and identified genes that need to be repressed to allow for the process.
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Project description:Grainyhead genes are involved in wound healing and developmental neural tube closure. In light of the high degree of similarity between the epithelial-mesenchymal transitions (EMTs) occurring in wound healing processes and the cancer stem cell-like compartment of tumors, including TGF-β-dependence, we investigated the role of a Grainyhead gene (GRHL2) in oncogenic EMT. Grainyhead was specifically down-regulated in the claudin-low subclass of mammary tumors and in the basal-B subclass of breast cancer cell lines. Functionally, GRHL2 suppressed TGF-β-induced, Twist-induced or spontaneous EMT, enhanced anoikis-sensitivity, and suppressed mammosphere generation in mammary epithelial cells. These effects were mediated, in part, by its suppression of ZEB1 expression, through direct repression of the ZEB1 promoter. GRHL2 also inhibited Smad-mediated transcription, and up-regulated mir200b/c as well as the TGF-β receptor antagonist, BMP2. The expression of GRHL2 in the breast cancer cell line MDA-MB-231 triggered a mesenchymal-to-epithelial transition and sensitized the cells to anoikis. These results indicate that GRHL2 is a suppressor of the oncogenic EMT. 3 biologic replicates for each cell line. Comparison of HMLE+Twist-ER cells expressing GRHL2/pMIG vs. HMLE+Twist-ER cells expressing empty pMIG.
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