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Expression of dioxin-responsive genes in human endometrial cells in culture
1School of Medicine, Catholic University of Taegu-Hyosung, Taegu, Republic of Korea. yangjh@cuth.cataegu.ac.kr
Abstract:
To investigate expression of dioxin-responsive genes in human endometrial cells with exposure to 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD), human endometrial stromal cells immortalized with temperature-sensitive SV40 T antigen were used for the experiments. Cells were treated with 0.1% DMSO or 0.1, 1, 10, or 100 nM TCDD for 24 h. Induction of interleukin-1beta (IL-1beta) and plasminogen activator inhibitor-2 (PAI-2) mRNAs was analyzed by reverse-transcription polymerase chain reaction. Expression of IL-1beta or PAI-2 mRNA in response to TCDD was increased in a dose-dependent fashion. The maximum increases of PAI-2 and IL-1beta mRNAs were observed at 100 and 10 nM TCDD, respectively. While cycloheximide treatment did not show a significant difference of PAI-2 mRNA levels between control and TCDD-treated cells, mRNA stability assay using actinomycin D showed that PAI-2 mRNA in TCDD-treated cells was about twofold more stable than the control cells. While expression of CYP1A1 mRNA was not detected and levels of ARNT mRNA were not altered by TCDD exposure, the amount of AhR mRNA was decreased dose dependently. The present study represents an initial attempt to determine the responses of dioxin-responsive genes in human endometrial cells following TCDD exposure. The results demonstrated that IL-1beta and PAI-2 genes are induced dose dependently in human endometrial cells with exposure to TCDD and expression of PAI-2 mRNA is controlled at the posttranscriptional level.
Insights
Exposure to dioxin (TCDD) increases expression of IL-1beta and PAI-2 genes in human endometrial cells. PAI-2 mRNA stability is enhanced post-transcriptionally, indicating a dose-dependent response to TCDD.
Area of Science:
- Environmental Toxicology
- Molecular Biology
- Reproductive Biology
Background:
- Dioxins, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), are environmental pollutants with known toxic effects.
- The impact of TCDD on dioxin-responsive genes in human endometrial cells is not well understood.
Purpose of the Study:
- To investigate the expression of dioxin-responsive genes in human endometrial cells following TCDD exposure.
- To determine the dose-dependent effects of TCDD on specific gene expression and mRNA stability.
Main Methods:
- Human endometrial stromal cells were treated with varying concentrations of TCDD (0.1-100 nM) for 24 hours.
- Reverse-transcription polymerase chain reaction (RT-PCR) was used to analyze the mRNA levels of IL-1beta, PAI-2, CYP1A1, ARNT, and AhR.
- mRNA stability assays were performed using actinomycin D.
Main Results:
- TCDD exposure induced a dose-dependent increase in IL-1beta and PAI-2 mRNA expression.
- PAI-2 mRNA stability was approximately doubled in TCDD-treated cells compared to controls.
- AhR mRNA levels decreased dose dependently, while CYP1A1 and ARNT mRNA levels remained unchanged or unaltered.
Conclusions:
- IL-1beta and PAI-2 are dioxin-responsive genes in human endometrial cells, with expression induced by TCDD in a dose-dependent manner.
- The expression of PAI-2 mRNA is regulated at the post-transcriptional level, involving enhanced mRNA stability.
- This study provides initial insights into the molecular responses of human endometrial cells to TCDD exposure.
