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Published on: May 23, 2014
Regulation of interleukin-8 gene expression in human endometrial cells in culture
A Arici1, J R Head, P C MacDonald
1Ceil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center at Dallas 75235-9051.
Insights
Human endometrial stromal and epithelial cells regulate interleukin-8 (IL-8) gene expression differently. Stromal cells increase IL-8 production upon stimulation, while epithelial cells maintain higher baseline levels and show distinct responses to IL-1 and serum.
Area of Science:
- Reproductive Biology
- Immunology
- Molecular Biology
Background:
- Interleukin-8 (IL-8) plays a crucial role in leukocyte recruitment to the endometrium.
- Understanding the differential regulation of IL-8 by endometrial stromal and epithelial cells is key to comprehending endometrial immune responses.
Purpose of the Study:
- To investigate the regulation of IL-8 gene expression in human endometrial stromal and epithelial cells.
- To elucidate the distinct mechanisms by which these cell types control IL-8 production.
Main Methods:
- Utilized well-characterized primary cultures of human endometrial stromal cells and glandular epithelial cells.
- Quantified IL-8 mRNA levels and protein accumulation in response to various stimuli including IL-1 alpha, tumor necrosis factor-alpha, and serum.
- Assessed IL-8 gene transcription and mRNA half-life in stromal cells.
Main Results:
- Endometrial stromal cells showed time- and concentration-dependent increases in IL-8 mRNA and protein upon stimulation with IL-1 alpha, TNF-alpha, or serum.
- Serum treatment in stromal cells modestly stimulated IL-8 gene transcription and significantly prolonged IL-8 mRNA half-life (>2.5-fold).
- Endometrial epithelial cells exhibited higher basal IL-8 mRNA levels and distinct regulatory responses to IL-1 and serum compared to stromal cells.
Conclusions:
- Endometrial stromal and epithelial cells possess unique regulatory pathways for IL-8 gene expression.
- Stromal cells primarily regulate IL-8 through transcriptional activation and mRNA stabilization, while epithelial cells maintain higher constitutive expression and respond differently to inflammatory signals.
Abstract:
In this study, we investigated the regulation of interleukin-8 (IL-8) gene expression in separated endometrial stromal and epithelial cells of human endometrium. This research was conducted as part of an analysis of the role of these cells in regulating the recruitment of leukocytes to the endometrium. Well-characterized model systems were used to study the regulation of endometrial IL-8 gene expression, namely, stromal cells in monolayer culture after first passage and glandular epithelium in primary culture. The levels of IL-8 mRNA and the accumulation of immunoreactive IL-8 in the medium of endometrial stromal cells is culture increased in a time- and concentration-dependent manner upon treatment with IL-1 alpha, tumor necrosis factor-alpha, or serum. The effects of IL-1 alpha plus serum on IL-8 mRNA levels were at least additive. Serum treatment caused a modest stimulation of IL-8 gene transcription (evaluated after 6 h of treatment) in endometrial stromal cells, but serum also acted in these stromal cells to prolong the half-life of IL-8 mRNA by more than 2.5-fold. The regulation of the levels of IL-8 mRNA in endometrial epithelial cells is distinctly different from that in stroma. First, the levels of IL-8 mRNA in non-treated epithelial cells in serum-free medium were much greater than those in stromal cells under similar conditions. Second, whereas the levels of IL-8 mRNA in endometrial epithelial cells also increased in response to serum and to IL-1 in the absence of serum, in the presence of serum, IL-1 treatment caused no appreciable change in the levels of IL-8 mRNA as was the case in endometrial stromal cells.
