| MST1重组腺病毒的制备及对人子宫内膜间质细胞蜕膜化的作用 |
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| 引用本文： | 赵静,华美娟,蒋玥,甄鑫,颜桂军,孙海翔.MST1重组腺病毒的制备及对人子宫内膜间质细胞蜕膜化的作用[J].生殖与避孕,2010,30(4):217-222,268. |
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| 作者姓名： | 赵静 华美娟 蒋玥 甄鑫 颜桂军 孙海翔 |
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| 作者单位： | 1. 南京师范大学生命科学学院江苏省分子医学生物技术重点实验室,南京,210046;南京大学医学院附属鼓楼医院生殖医学中心,南京,210008
2. 南京大学医学院附属鼓楼医院生殖医学中心,南京,210008 |
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| 基金项目： | 国家自然科学基金(30900727); 江苏省自然科学基金(BK2009038); 南京市卫生局资金(QYK09172)资助项目 |
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| 摘 要： | 目的:探讨丝/苏氨酸激酶MST1在人子宫内膜间质细胞蜕膜化过程中的作用。方法:制备Ad-FLAG-MST1重组腺病毒,用于感染人子宫内膜间质细胞;采用荧光定量PCR实验结合腺病毒介导的MST1基因过表达,分析人子宫内膜间质细胞蜕膜化进程中MST1的表达及其对蜕膜化特异性基因的调控。结果:获得滴度为2×1011ifu/ml的重组Ad-FLAG-MST1腺病毒,该腺病毒感染人子宫内膜间质细胞后,可高水平表达FLAG-MST1融合蛋白,过表达的MST1显著增加人子宫内膜间质细胞中PRL、IGFBP1、TIMP3和DCN等蜕膜化特异性基因的表达;体外培养的人子宫内膜间质细胞经8-Br-cAMP(0.5mmol/L)和MPA(1μmol/L)刺激后,细胞中MST1总蛋白水平增加3倍以上,同时磷酸化的MST1(Thr183)显著增加。结论:MST1的活化可能参与子宫内膜间质细胞的蜕膜化相关基因的调控。
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| 关 键 词： | MST1 人子宫内膜间质细胞 蜕膜化 |
Evaluation of Hysteroscopic-guided Removal of Intrauterine Device in Preparation of Recombinant Adenovirus Harboring MST1 and Its Role in Decidualization of Human Endometrial Stromal Cells |
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| Jing ZHAO,Mei-juan HUA,Yue JIANG,Xin ZHEN,Gui-jun YAN,Hai-xiang SUN.Evaluation of Hysteroscopic-guided Removal of Intrauterine Device in Preparation of Recombinant Adenovirus Harboring MST1 and Its Role in Decidualization of Human Endometrial Stromal Cells[J].Reproduction and Contraception,2010,30(4):217-222,268. |
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| Authors: | Jing ZHAO Mei-juan HUA Yue JIANG Xin ZHEN Gui-jun YAN Hai-xiang SUN |
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| Institution: | Jing ZHAO1,2,Mei-juan HUA2,Yue JIANG2,Xin ZHEN2,Gui-jun YAN2,Hai-xiang SUN2 (1. Jiangsu Key Laboratory for Molecular , Medical Biotechnology,School of Life Sciences,Nanjing Normal University,Nanjing,210097) (2. Reproductive Medicine Center,the Affiliated Drum Tower Hospital of Nanjing University Medical School,210008) |
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| Abstract: | Objective: To investigate the effects of MST1 kinase during the decidualization process of human endometrial stromal cells. Methods: Construct adenovirus-FLAG-MST1 to infect human endometrial stromal cells. Real-time quantitative-PCR and adenovirus-mediated overexpression of MST1 were performed to study the regulation role of MST1 in the specific genes of decidualization during the decidualization process.Results: A total of 2×10~(11) ifu/ml adenovirus-FLAG-MST1 was obtained, which can continue overexpressing FLAG-MST1 fusion protein in human endometrial stromal cells. MST1 overexpression resulted in an increase of the expression of PRL, IGFBP1, TIMP3 and DCN in human endometrial stromal cells. When treated with 0.5 mmol/L 8-Br-cAMP plus 1 μmol/L medroxyprogesterone, the total MST1 level was similar to untreated cells, whereas the levels of phosphorylated MST1 (phospho-MST1) were markedly increased. Conclusion: MST1 is a novel kinease for decidualization in human endometrial stromal cells. |
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| Keywords: | MST1 human endometrial stromal cells decidualization |
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