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CircRNA-sequencing analysis of Ishikawa human endometrial carcinoma cells treated with MPA or not

ABSTRACT: circRNA-sequencing analysis of Ishikawa human endometrial carcinoma cells treated with MPA or not

PROVIDER: PRJNA748159 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15195244_1.fastq.gz	Fastqsanger.gz
	SRR15195244_2.fastq.gz	Fastqsanger.gz
	SRR15195245_1.fastq.gz	Fastqsanger.gz
	SRR15195245_2.fastq.gz	Fastqsanger.gz
	SRR15195246_1.fastq.gz	Fastqsanger.gz

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Project description:Every year more than 42,000 women die of endometrial cancer, mainly due to recurrent or metastatic disease. The presence of tumor infiltrating lymphocytes (TILs) as well as progesterone receptor (PR) positivity has been correlated with improved prognosis. This study describes two mechanisms by which progesterone inhibits metastatic spread of endometrial cancer: by stimulating T-cell infiltration and by inhibiting epithelial-to-mesenchymal cell transition (EMT). Paraffin sections from patients with (n=9) or without (n=10) progressive endometrial cancer (recurrent or metastatic disease) were assessed for the presence of CD4+ (helper), CD8+ (cytotoxic) and Foxp3+ (regulatory) T-lymphocytes and PR expression. Progressive disease was observed to be associated with significant loss of TILs and loss of PR expression. Frozen tumor samples, used for genome-wide expression analysis, showed significant regulation of pathways involved in immunosurveillance, EMT and metastasis. For a number of genes, such as CXCL14, DKK1, DKK4 and WIF1, quantitive RT-PCR was performed to verify down regulation in progressive disease. To corroborate the role of progesterone in regulating invasion, Ishikawa (IK) endometrial cancer cell lines stably transfected with PRA (IKPRA), PRB (IKPRB) and PRA+PRB (IKPRAB) were cultured in the presence/absence of progesterone (MPA) and used for genome-wide expression analysis, Boyden- and wound healing migration assays, and IHC for known EMT makers. IKPRB and IKPRAB cell lines showed MPA induced inhibition of migration and loss of the mesenchymal marker vimentin at the invasive front of the wound healing assay. Furthermore, pathway analysis of significantly MPA-regulated genes showed significant down regulation of important pathways involved in EMT, immunesuppression and metastasis: such as IL6-, TGF-β and Wnt/β-catenin signalling. Intact progesterone signaling in non-progressive endometrial cancer seems to be an important factor stimulating immunosurveillance and inhibiting transition from an epithelial to a more mesenchymal, more invasive phenotype. The PRA- and PRB-expressing Ishikawa endometrial cancer cell line (IKPRAB36) was cultured for 48h in the absence or presence of 1nM MPA (n = 3). The RNA was isolated and used for hybridization on Affymetrix microarrays. Cells cultured in the presence of MPA were compared with cells cultured in the absence of MPA.

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CircRNA-sequencing analysis of Ishikawa human endometrial carcinoma cells treated with MPA or not

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