Project description:Expression profiles in decidualized and non-decidualized endometriotic cyst stromal cells (ECSCs) and normal endometrial stromal cells (NESCs)
Project description:Endometriotic cyst stromal cells (ECSCs) were isolated from ovarian endometriotic cyst and cultured for 48 hours in the floating three-dimensional collagen gel culture or in the conventional two-dimensional culture. Total mRNAs were extracted and subjected to gene expression microarray.
Project description:Accumulating evidences suggest that various epigenetic aberrations play definite roles in the pathogenesis of endometriosis. The aim of this study is to identify 1) the panel of aberrantly expressed genes that are epigenetically suppressed by histone acetylation in endometriosis; 2) the roles of CCAAT/enhancer-binding protein (C/EBP) alpha, one of the candidate molecules whose expression was epigenetically repressed in endometriotic cyst stromal cells (ECSCs), in the pathogenesis of endometriosis; and 3) the efficacy of the histone deacetylase inhibitors for the treatment of endometriosis. Subconfluent ECSCs cultured in 10-cm dish were further incubated for 72 h with or without VPA (8 mM) and/or 5aza. Total RNA from untreated ECSCs (n=4), VPA-treated ECSCs (n=4), 5aza-treated ECSCs (n=4), and VPA- and 5aza-treated ECSCs (n=4) was extracted and subjected to gene expression microarray analysis.
Project description:To identify the miRNAs which regulate the decidualization of ECSCs, we explored differentially expressed miRNAs between untreated ECSCs (n=4) and decidualized ECSCs (n=4) using miRNA microarray. The miRNA expression profiles in untreated ECSCs and decidualized ECSCs were obtained, then the miRNAs were filtered by signal intensity (50.0<, at least one out of 8 samples have values within range) and flags (at least 17% of cases have good flags). We established the criteria for regulated genes: ratio ≥2.0-fold for up-regulated genes and ratio ≤0.5 for down-regulated genes.
Project description:We previously demonstrated that the expression of histone deacetylase (HDAC) 10 is elevated in ovarian endometriotic cyst stromal cells (ECSCs) compared to normal endometrial stromal cells (NESCs). HDAC10 inhibitor, tucidinostat (10 μM) inhibited the cell proliferation of ECSCs and induced the apoptosis and G0/G1 cell cycle arrest of these cells. GThe objective of this study to unveil the role of HDAC10 in the pathogenesis of endometriosis. We cultured ECSCs with an HDAC10 inhibitor, tucidinostat (10 μM), isolated the total RNAs, and subjected to next-generation RNA sequencing to identify the target genes of HDAC10. Gene Ontology analysis identified upregulated genes. We will introduce these genes into endometriosis stromal cells and examine whether they induce cell proliferation inhibition and apoptosis promotion similar to HDAC10 inhibitors. This will help us evaluate whether HDAC10 inhibitors are promising candidates for endometriosis treatment. We found that somatostatin receptor 2 (SSTR2), dachshund homolog 11 (DACH1), and interferon regulatory factor 6 (IRF6) are the possible target genes of HDAC10.
Project description:Accumulating evidences suggest that various epigenetic aberrations play definite roles in the pathogenesis of endometriosis. The aim of this study is to identify 1) the panel of aberrantly expressed genes that are epigenetically suppressed by histone acetylation in endometriosis; 2) the roles of CCAAT/enhancer-binding protein (C/EBP) alpha, one of the candidate molecules whose expression was epigenetically repressed in endometriotic cyst stromal cells (ECSCs), in the pathogenesis of endometriosis; and 3) the efficacy of the histone deacetylase inhibitors for the treatment of endometriosis.
Project description:To identify the mRNAs which regulate the decidualization of ECSCs, we explored differentially expressed mRNAs between untreated ECSCs (n=4) and decidualized ECSCs (n=4) using gene expression microarray. The mRNA expression profiles in untreated ECSCs and decidualized ECSCs were obtained, then the mRNAs were filtered by signal intensity (50.0<, at least one out of 8 samples have values within range) and flags (at least 58% of cases have good flags). We established the criteria for regulated genes: Z-score ≥2.0 and ratio ≥2.0-fold for up-regulated genes, and Z-score ≤–2.0 and ratio ≤0.5 for down-regulated genes.
