Quantitative and qualitative analysis of estrogen receptor mRNAs on peripheral mononuclear blood cells from females in different estrogen-endocrine circumstances

不同雌激素内分泌环境下女性外周血单个核细胞雌激素受体mRNA的定量和定性分析

• 批准号: 12671623
• 负责人: HONJO Hideo
• 金额: $ 2.05万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671623/
• 关键词: HRT; Peripheral mononuclear cell; estrogen receptor; エストロゲン受容体; リンパ球

Object:The expression of estrogen receptor (ER) on peripheral mononuclear blood cells (PMBC) was reported. Recent progress in the study of ER revealed the variety of forms and indicated the related functions. But till now, no detailed examination has been made about the expression of ERs and peripheral mononuclear blood cell (PMBC). To see the mechanism of hormone replacement therapy (HRT) through the PMBC, this study was designed to make a quantitative and qualitative analysis about the variant kinds of ERs on mRNA level on PMBC from the women in variant endocrine circumstances.Material and Method:With informed consent,2 ml of peripheral whole blood was obtained in EDTA tube from women with regular menstrual cycle (MC ; n=42), menopausal women without HRT (MP ; n40), women with HRT (HR ; n=42) and pregnant women (PR ; n=44). PMBC was isolated and total RNA was extracted. Quantitative RT-PCR was done about Wild type ER α (WtER α), exon 3 alternatively spliced ERα (Δ3ERα),Δ4ERα,Δ5ERα,Δ7 … More ERα, WtERβ and Δ5ERβ mRNAs using specific primers. All data-was corrected with β actin as Internal control which was measured in the same sample simultaneously.Result:WtERα mRNA was detected in all samples. ERα mRNA is dominantly expressed in PMBC and its expression level is influenced with the systemic estrogen levels measured in peripheral blood serum. After menopause it decreased significantly but with 6 month HRT, it was recovered to the level of MC. WtERβ mRNA was detected from 39% to 79% of the cases in each group. But its expression level showed no correlation with individual estrogen level. Δ3ERα mRNA and Δ5ERα mRNA were rarely detected in PMBC. The expression of Δ4ERα mRNA and Δ7ERα mRNA tended to decrease after menopause, but it showed no correlation with serum estrogen level and showed no change with HRT.Conclusion:In PMBC, WtERα mRNA is dominantly expressed and it is supposed that serum estrogen level exerted great influence to the expression level. After menopause, it decreases but HRT of more than 6 month can work effectively to recover it to the level of MC, but could not recover the expression level of ΔERα mRNAs. Less

目的：研究外周血单个核细胞（PMBC）雌激素受体（ER）的表达。近年来ER的研究进展揭示了ER的多种形式及其相关功能。但迄今为止，对ER和外周血单个核细胞（PMBC）的表达还没有详细的研究。为探讨激素替代疗法（hormone replacement therapy，HRT）的作用机制，本研究对不同内分泌状态的妇女外周血单个核细胞（PMBC）上不同种类的雌激素受体（ER）mRNA进行定量和定性分析（MC ; n=42）、未接受HRT的绝经妇女（MP ; n = 40）、接受HRT的妇女（HR ; n=42）和妊娠妇女（PR ; n=44）。分离PMBC并提取总RNA。对野生型ER α（WtER α）、外显子3可变剪接ERα（Δ3ERα）、Δ 4 ER α、Δ5ERα、Δ7 ...更多信息 ERα、WtERβ和Δ5ERβ mRNA。结果：所有标本中均检测到WtERα mRNA。ERα mRNA主要在PMBC中表达，其表达水平受外周血雌激素水平的影响。绝经后明显下降，经6个月HRT后恢复至MC水平。WtERβ mRNA的表达率为39%~ 79%。但其表达水平与个体雌激素水平无关。Δ 3 ER α mRNA和Δ 5 ER α mRNA在PMBC中很少表达。绝经后Δ 4 ER α mRNA和Δ 7 ER α mRNA表达呈下降趋势，但与血清雌激素水平无相关性，与HRT无相关性。结论：PMBC中WtERα mRNA以优势表达为主，血清雌激素水平对其表达水平有较大影响。绝经后，Δ ER α mRNA的表达水平下降，但6个月以上的HRT能有效恢复到MC水平，但不能恢复ΔERα mRNA的表达水平。少