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雌激素受体α抑制剂MPP在小鼠囊胚形成和滋养层干细胞中影响YAP核定位

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雌激素受体α抑制剂MPP在小鼠囊胚形成和滋养层干细胞中影响YAP核定位
MPP, inhibitor of estrogen receptor α, affects YAP nuclear localization during mouse blastocyst formation and in Trophoblast Stem Cells
上传时间:2019/4/18 15:03:08    作者:许颂华,朱灵华,刘玥,王晓江,钟玉环,苏杨,许丽旋,王世鄂|下载

许颂华1, 2,朱灵华3,刘玥1, 2,王晓江2,钟玉环2,苏杨2,许丽旋2,王世鄂1, 2 *
(1干细胞工程与再生医学福建省高校重点实验室,福州 350108;2福建医科大学基础医学院人体解剖学与组织胚胎学系,福州  350108;3福建医科大学基础医学专业2012级本科班,福州  350108)

Xu Songhua1, 2, Zhu Linghua3, Liu Yue1, 2, Wang Xiaojiang2, Zhong Yuhuan2, Su Yang2, Xu Lixuan2, Wang Shie1, 2 *
(1Key Laboratory of Stem Cell Engineering and Regenerative Medicine of Fujian Provincial Colleges and Universities; 2Department of Human Anatomy and Histo-Embryology, School of Basic Medical Sciences; 3Undergraduate Class recruited in 2012 of Basic Medicine, Fujian Medical University, Fuzhou 350108, China)
摘要:目的 研究雌激素受体α(estrogen receptor α,ERα)抑制剂甲基-哌啶-吡唑(methyl-piperidino-pyrazole,MPP)在小鼠桑葚胚和滋养层干细胞(trophoblast stem cells,TSCs)中对YAP的影响。方法  收集昆明(kunming,KM)小鼠8-细胞胚,置于0μmol/L(对照组)和5μmol/L(实验组)MPP中培养,分别于8h、12h和24h后收集各组桑葚胚,采用免疫荧光技术观察YAP表达,采用Real Time-PCR检测MPP处理24h后Yap mRNA表达变化;将小鼠TSCs置于0μmol/L、2.5μmol/L、5μmol/L和10μmol/L MPP中培养,48h后观察细胞形态改变,分别检测Sox2、YAP、Cdx2 mRNA和蛋白表达水平。结果 小鼠8-细胞胚经MPP处理24h后,桑葚胚YAP蛋白核定位水平降低,Yap mRNA水平无显著改变;经MPP处理48h后,5μmol/L组小鼠TSCs细胞出现细胞团块,10μmol/L组细胞增殖明显受抑制;与对照组相比,5μmol/L组细胞Sox2 mRNA表达水平升高,Yap和Cdx2 mRNA水平无显著改变,团块状细胞中的YAP蛋白失去明显的核内定位,SOX2和CDX2阳性细胞的表达更加密集。结论  ERα在小鼠桑葚胚和TSCs中调控YAP核定位,其在TSCs细胞中的作用与CDX2和SOX2的表达有关。
关键词:雌激素受体α;桑葚胚;滋养层干细胞;YAP
〔中图分类号〕R321             〔文献标识码〕A             DOI:10.16705/ j. cnki. 1004-1850. 2019. 01. 002 
Abstract: Objective To study the effects of Estrogen Receptor alpha (ERα) inhibitor methyl-piperidino-pyrazole (MPP) on YAP localization in mouse morula and Trophoblast Stem cells (TSCs). Methods  8-cell embryos of KM mice were collected and cultured in 0 μmol/L (control group) and 5 μmol/L (experimental group) MPP, respectively. The morulae of each group were collected after 8h, 12h and 24h culture, respectively. The expression of YAP was observed by immunofluorescence, and the expression of Yap mRNA was detected by Real Time-PCR after MPP treatment for 24 hours. The TSCs were cultured in 0 μmol/L, 2.5 μmol/L, 5 μmol/L and 10 μmol/L MPP. After 48 h, TSCs morphology was observed and their Sox2, Yap, Cdx2 mRNA and protein expression levels were detected. Results After treatment with MPP for 24 h, the nuclear localization level of YAP protein in morula stage embryos decreased, while the level of Yap mRNA did not change significantly. After 48 h treatment with MPP, cell clumps appeared in TSCs cells of 5 μmol/L group, while the proliferation of cells in the 10 μmol/L group was significantly inhibited. Compared with the control group, the expression of Sox2 mRNA was increased in the 5 μmol/L group, however, the levels of Yap and Cdx2 mRNA didn’t significantly change. The YAP protein in the cell clumps lost obvious nuclear localization and SOX2 and CDX2 positive cells were more densely expressed. Conclusion  ERα regulates YAP nuclear localization in mouse morula and TSCs, and its role in TSCs is related to the expression of CDX2 and SOX2.
Keywords:Estrogen receptor α; morula; trophoblast stem cells; YAP

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