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Characterizing EMT-Linked Heterogeneity in Breast Cancer Circulating Tumor Cells at the Single-Cell Level

ABSTRACT: Characterizing EMT-Linked Heterogeneity in Breast Cancer Circulating Tumor Cells at the Single-Cell Level

PROVIDER: PRJNA1115116 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29142600_1.fastq.gz	Fastqsanger.gz
	SRR29142600_2.fastq.gz	Fastqsanger.gz
	SRR29142601_1.fastq.gz	Fastqsanger.gz
	SRR29142601_2.fastq.gz	Fastqsanger.gz
	SRR29142602_1.fastq.gz	Fastqsanger.gz

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Project description:Cyclin D1 is a well characterised cell cycle regulator with established oncogenic capabilities. Despite these properties, studies report contrasting links to tumour aggressiveness. It has previously been shown that silencing cyclin D1 increases the migratory capacity of MDA-MB-231 breast cancer cells with concomitant increase in ‘inhibitor of differentiation 1’ (ID1) gene expression. Id1 is known to be associated with more invasive features of cancer and with the epithelial-mesenchymal transition (EMT). Here, we sought to determine if the increase in cell motility following cyclin D1 silencing was mediated by Id1 and enhanced EMT-features. To further substantiate these findings we aimed to delineate the link between CCND1, ID1 and EMT, as well as clinical properties in primary breast cancer. The increase in cell migration following cyclin D1 silencing in MDA-MB-231 cells was abolished by Id1 siRNA treatment and we observed cyclin D1 occupancy of the Id1 promoter region. Moreover, ID1 and SNAI2 gene expression was increased following cyclin D1 knock-down, an effect reversed with Id1 siRNA treatment. Similar migratory and SNAI2 increases were noted for the ER-positive ZR75-1 cell line, but in an Id1 independent manner. In a meta-analysis of 1107 breast cancer samples, CCND1 and ID1 gene expression were associated with mesenchymal-markers including SNAI1, SNAI2 and TWIST1, and with clinicopathological parameters. Finally, a greater percentage of CCND1low/ID1high tumours were found in the EMT-like ‘claudin-low’ subtype of breast cancer than in other subtypes. Together, these results indicate that increased migration of MDA-MB-231 cells following cyclin D1 silencing can be mediated by Id1 and is linked to an increase in EMT markers. Moreover, we have confirmed a relationship between cyclin D1, Id1 and EMT in primary breast cancer, supporting our in vitro findings that low cyclin D1 expression can be linked to aggressive features in subgroups of breast cancer. MDA-MB-231 cells were transfected with cyclin D1, CDK4/6 or control siRNA.

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Characterizing EMT-Linked Heterogeneity in Breast Cancer Circulating Tumor Cells at the Single-Cell Level

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