Project description:In ovarian endometrioma, much iron, which is derived from blood in cyst, deposit in stroma. This iron generates oxidative stress by Fenton reaction, which is supposed to affect endometriotic stromal cells. We used gene expression microarrays to find influence of oxidative stress on endometriotic stromal cells. Gene expression microarrays revealed no statistically differences caused by oxidative stress.
Project description:The development and progression of endometriotic lesions are poorly understood, but immune cell dysfunction and inflammation are closely associated with the pathophysiology of endometriosis. A lack of suitable 3D in vitro models permitting the study of interactions between cell types and the microenvironment is a contributing factor. To address this limitation, we developed endometriotic organoids (EO) to explore the role of epithelial-stromal interactions and model peritoneal cell invasion associated with lesion development. Using a non-adherent microwell culture system, spherical organoids were generated with endometriotic epithelial cells (12Z) combined with immortalized endometriotic stromal cells (iEc-ESC) or immortalized uterine stromal cells (iHUF). Organoids self-organized with stromal cells occupying the center and epithelial cells on the periphery of the organoid. Endometriotic organoids (EO), containing iEc-ESC, resulted in the development of stratified 12Z epithelial cells compared to those with iHUF where the 12Z cells developed as a single layered epithelium. Transcriptomic analysis found 4,522 differentially expressed genes (DEG) between EO and 12Z/iHUF organoids, and the top DEG included increased expression of interleukins and prostaglandin synthase enzymes. An overlap of the EO DEG with baboon endometriotic lesions was highly significant. Finally, to mimic invasion of endometrial tissue into the peritoneum, a model was developed using EO and extracellular matrix containing human peritoneal mesothelial cells (LP9). Invasion of EO into the extracellular matrix-LP9 layer was increased in presence of estrogen or THP1-derived proinflammatory macrophages. Taken together, our results strongly support the concept that EO are an appropriate model for dissecting mechanisms that contribute to endometriotic lesion development.
Project description:To identify specific markers of rectovaginal endometriotic nodule vasculature, highly enriched preparations of vascular endothelial cells and pericytes were obtained from endometriotic nodules and control endometrial and myometrial tissue by laser capture microdissection (LCM), and gene expression profiles were screened by microarray analysis. Of the 18,400 transcripts on the arrays, 734 were significantly overexpressed in vessels from fibromuscular tissue and 923 in vessels from stromal tissue of endometriotic nodules, compared to vessels dissected from control tissues. The most frequently expressed transcripts included known endothelial cell-associated genes, as well as transcripts with little or no previous association with vascular cells. The higher expression in blood vessels was further corroborated by immunohistochemical staining of 6 potential markers, 5 of which showed strong expression in pericytes. The most promising marker was matrix Gla protein, which was found to be present in both glandular epithelial cells and vascular endothelial cells of endometriotic lesions, while it was barely expressed at all in normal endometrium.
Project description:miRNA high-throughput sequencing was used to investigate endometriosis lesion-specific miRNA expression profiles by comparing a set of paired samples of peritoneal endometriotic lesions and matched healthy surrounding tissue together with eutopic endometrium of the same patients. We found that miRNAs of surrounding peritoneal tissue mask most of the miRNA expression differences that could originate from endometriotic tissue and thus only miRNAs with significantly different levels in the endometriotic lesions compared to peritoneal tissue were detected. According to the results of this study, two miRNAs – miR-34c and miR-449a showed remarkably higher expression in lesions compared to healthy tissue.
Project description:The aim of this study is to compare transcriptome profiling of EGR1 siRNA treated immortalized human endometriotic epithelial cells expression luciferase (iHEECs/Luc) by RNA-sequencing Methods: The mRNA profiles of non-targeting (NT) and EGR1 siRNA treated immortalized human endometriotic epithelial cells expression luciferase (iHEECs/Luc) were generated by deep sequencing, in triplicate using Illumina NovaSeq-6000 sequencers with 2×100 paired-end reads. Basecalls and demultiplexing were performed with Illumina’s RTA version 1.9 and bcl2fastq2 software with a maximum of one mismatch in the indexing read. RNA-seq reads were then aligned to the human genome (Genome Reference Consortium Human Build hg38) with STAR version 2.5.1a. qRT–PCR validation was performed using TaqMan assays. Results: Using a 1.5-fold cutoff and Benjamini-Hochberg false discovery rate (FDR) of <0.05 threshold for inclusion, we identified 76 differentially expressed genes (DEGs) between (NT) and EGR1 siRNA treated immortalized human endometriotic epithelial cells. Conclusions: Our study represents the first detailed analysis of EGR1 siRNA treated immortalized human endometriotic epithelial cells expression luciferase (iHEECs/Luc) transcriptomes, with biologic replicates, generated by RNA-seq technology.
Project description:It has been shown that the Chelidonium majus extract NSC-631570 has growth and invasion inhibiting properties on head and neck carcinoma cell lines. The Affymetrix PrimeView array was used to gain a first view on the influence of NSC-631570 on gene expression of head and neck cancer cells.