Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptomic effects of estradiol treatment on cultured human uterine smooth muscle cells

ABSTRACT: The myometrium is an important reproductive tissue composed primarily of smooth muscle cells. Contractility of the myometrial smooth muscle cells during the estrous cycle and pregnancy is modulated by estrogen. Despite much research, little is known about the molecular mechanism by which estrogen regulates myometrial contractility. This study investigates global gene expression profile of cultured human uterine smooth muscle cells (hUtSMCs) following 17M-NM-2-estradiol (E2) treatment using cDNA microarray technology. In response to E2 treatment 540 genes were identified as significantly differentially expressed. Gene ontology analysis identified several significant biological processes for these genes including muscle contraction, gland development, cell migration, cell adhesion, apoptosis, response to external stimulus and phosphorylation. In this study evidence is presented that in human smooth muscle cells 17M-NM-2-estradiol, contrary to expectations, altered expression of several genes that may favor myometrial relaxation. Genes related to focal adhesion function were downregulated suggesting a loosening of SMC connections to the ECM while upregulation of MMP9 is also associated with weakened ECM interactions. The hUtSMC system is an additional useful model to investigate steroid effects on smooth muscle cells in isolation from other myometrial cell types. In this study human myometrial smooth muscle cells (hUtSMC) purchased from Lonza were cultured and treated with 17beta estradiol. RNA was isolated after 6h, 24h and 72h from both treated and untreated control cells. The experiment was repeated at least 3 times.

ORGANISM(S): Homo sapiens

SUBMITTER: Michael Cairns

PROVIDER: E-GEOD-51470 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The myometrium is an important reproductive tissue composed primarily of smooth muscle cells. Contractility of the myometrial smooth muscle cells during pregnancy and labour is modulated by hormones. Despite much research, little is known about the molecular mechanism by which estrogen and progesterone regulate myometrial contractility. This study investigates global gene expression profile of cultured human uterine smooth muscle cells (hUtSMCs) following 17Î²-estradiol (E2) and/or progesterone treatments using cDNA microarray technology. Furthermore the effects of addition of the progesterone inhibitor RU486 were investigated. Many genes were regulated in the presence of P4 or E2 alone but almost six times these numbers were regulated in the presence of the combination. The majority of these genes returned to near baseline levels (control samples) on addition of RU486. In total 796 annotated genes were significantly differentially expressed in the combined presence of P4 and E2 (relative to untreated levels). Furthermore, 666 annotated genes were significantly regulated by the addition of RU486 to the P4/E2 combination (relative to P4/E2 levels). Gene ontology analysis of these differentially expressed genes revealed a strong association of P4/E2 treatment with the downregulated expression of genes involved in cell communication, signal transduction, channel activity, inflammatory response and differentiation. Upregulated processes included cell survival (anti-apoptosis), gene transcription, steroid hormone biosynthesis, muscle development, insulin receptor signaling and cell growth. Functional withdrawal of progesterone using RU486 effectively reversed the processes induced by P4/E2 treatment. The hUtSMC system is an additional useful model to investigate steroid effects on smooth muscle cells in isolation from other myometrial cell types. In this study human myometrial smooth muscle cells (hUtSMCs) purchased from Lonza were cultured and treated with 17beta estradiol and progesterone either alone or in combination. In addition the 17beta estradiol and progesterone combined treated samples were also treated with the progesterone inhibitor RU486 (mifepristone). RNA was isolated after 72h from both treated and untreated control cells. The experiment was repeated 3 times. Please note that slides were scanned three times successively (i.e. 3 .gpr files per sample) and the median raw values were used as input raw data for GeneSpring analysis (Median_Table_InputRAW.xlsx).

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Transcriptomic effects of estradiol treatment on cultured human uterine smooth muscle cells

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