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培养的人子宫内膜细胞中二噁英反应基因的表达。

Expression of dioxin-responsive genes in human endometrial cells in culture.

作者信息

Yang J H

机构信息

School of Medicine, Catholic University of Taegu-Hyosung, Taegu, Republic of Korea.

出版信息

Biochem Biophys Res Commun. 1999 Apr 13;257(2):259-63. doi: 10.1006/bbrc.1999.0451.

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.

摘要

为了研究暴露于2,3,7,8-四氯二苯并对二恶英(TCDD)的人子宫内膜细胞中二恶英反应基因的表达,使用对温度敏感的SV40 T抗原永生化的人子宫内膜基质细胞进行实验。细胞用0.1%二甲亚砜(DMSO)或0.1、1、10或100 nM TCDD处理24小时。通过逆转录聚合酶链反应分析白细胞介素-1β(IL-1β)和纤溶酶原激活物抑制剂-2(PAI-2)mRNA的诱导情况。TCDD诱导的IL-1β或PAI-2 mRNA表达呈剂量依赖性增加。PAI-2和IL-1β mRNA的最大增加分别在100 nM和10 nM TCDD时观察到。虽然环己酰亚胺处理未显示对照细胞和TCDD处理细胞之间PAI-2 mRNA水平有显著差异,但使用放线菌素D的mRNA稳定性分析表明,TCDD处理细胞中的PAI-2 mRNA比对照细胞稳定约两倍。虽然未检测到CYP1A1 mRNA的表达,且TCDD暴露未改变芳烃受体核转运蛋白(ARNT)mRNA的水平,但芳烃受体(AhR)mRNA的量呈剂量依赖性降低。本研究是确定TCDD暴露后人子宫内膜细胞中二恶英反应基因反应的初步尝试。结果表明,暴露于TCDD的人子宫内膜细胞中,IL-1β和PAI-2基因呈剂量依赖性诱导,且PAI-2 mRNA的表达在转录后水平受到调控。

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