本文選題：囊胚　切入點：極滋養(yǎng)層細胞　出處：《北京大學(xué)學(xué)報(醫(yī)學(xué)版)》2017年06期 　論文類型：期刊論文

【摘要】：目的:研究人胚胎植入前囊胚滋養(yǎng)外胚層細胞基因的空間表達。方法:輔助生殖來源人受精后第6天Gardner評分5AA的囊胚,顯微鏡下微吸管機械分離滋養(yǎng)外胚層細胞,對囊胚滋養(yǎng)外胚層極滋養(yǎng)層細胞和壁滋養(yǎng)層細胞采用單細胞測序檢測,選取經(jīng)t檢驗計算所得P0.05且基因表達差異≥2倍的差異表達基因,采用生物信息學(xué)軟件對篩選的差異基因進行無監(jiān)督層次聚類分析和基因本體(gene ontology,GO)功能分類分析,對差異表達基因進行注釋和生物功能富集,并用全基因組對差異基因功能進一步注釋。結(jié)果:對2枚受精后第6天的5AA囊胚滋養(yǎng)外胚層8個極滋養(yǎng)層細胞和8個壁滋養(yǎng)層細胞測序,發(fā)現(xiàn)極滋養(yǎng)層細胞306個基因上調(diào),壁滋養(yǎng)層細胞75個基因上調(diào)。無監(jiān)督聚類分析結(jié)果顯示外胚層細胞分成極滋養(yǎng)層細胞和壁滋養(yǎng)層細胞兩群,屬于兩種不同類型和生物功能的細胞群。差異基因功能注釋顯示,極滋養(yǎng)層細胞上調(diào)基因GO生物學(xué)功能主要是轉(zhuǎn)錄、能量代謝、蛋白合成、轉(zhuǎn)運、氧化應(yīng)激、離子轉(zhuǎn)運、蛋白合成及運輸、細胞周期調(diào)節(jié)、肌動蛋白增長等,主要參與泛素介導(dǎo)的蛋白水解、氧化磷酸化、Wnt信號通路、雌雄激素代謝等信號通路;壁滋養(yǎng)層細胞上調(diào)基因GO生物學(xué)功能主要是蛋白分解代謝、細胞周期停滯、細胞凋亡、激活絲裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信號通路、碳水化合物運輸、突觸負(fù)調(diào)節(jié)、細胞生長、鈣通道激活、正向B細胞分化、T細胞凋亡等,主要參與B細胞受體、T細胞受體、白細胞跨內(nèi)皮移植、血管內(nèi)皮生長因子(vascular endothelial growth factor,VEGF)表達、間隙連接、促性腺激素釋放激素(gonadotropin-releasing hormone,GnRH)分泌、細胞凋亡等信號通路。結(jié)論:從空間維度揭示囊胚極/壁滋養(yǎng)層細胞的基因表達,顯示胚胎植入前囊胚極/壁滋養(yǎng)外胚層相互協(xié)調(diào),精細調(diào)節(jié)囊胚孵化和胚胎植入過程,進一步數(shù)據(jù)分析有望尋找到調(diào)控胚胎植入的內(nèi)源性特異分子。
[Abstract]:Objective: to study the spatial expression of human blastocyst trophodermal cell gene in preimplantation human blastocyst. Methods: the blastocyst with Gardner score of 5AA on the 6th day after assisted reproduction was used to isolate the trophoblast ectodermal cells by micropipette mechanically. Single cell sequencing was used to detect trophoblast cells and parietal trophoblast cells of blastocyst trophoderm. Bioinformatics software was used for unsupervised hierarchical cluster analysis and functional classification analysis of gene ontologyGO. the differentially expressed genes were annotated and biofunctional enrichment was carried out. Results: on the 6th day after fertilization, the 5AA blastocysts were sequenced from 8 extremely trophoblast cells and 8 parietal trophoblast cells, and 306 genes were up-regulated in the very trophoblast cells. The results of unsupervised cluster analysis showed that the ectodermal cells were divided into two groups of extremely trophoblastic cells and parietal trophoblastic cells, which belonged to two different types and biological function groups. The up-regulation of go gene in trophoblast cells mainly includes transcription, energy metabolism, protein synthesis, transport, oxidative stress, ion transport, protein synthesis and transport, cell cycle regulation, actin growth and so on. Ubiquitin mediated protein hydrolysis, oxidative phosphorylation of Wnt signaling pathway, androgen metabolism, etc. The up-regulated gene go biological function of trophoblastic cells is mainly protein catabolism, cell cycle arrest, cell apoptosis, etc. Activation of mitogen-activated protein kinases (MAPK) signaling pathway, carbohydrate transport, synaptic negative regulation, cell growth, calcium channel activation, positive differentiation of T cells into B cells, apoptosis of T cells, etc. The expression of vascular endothelial growth factor (VEGF), gap junction, gonadotropin-releasing hormone gonadotropin-releasing hormone (GnRH), vascular endothelial growth factor (VEGF), vascular endothelial growth factor (VEGF) and gonadotropin-releasing hormone (GnRH) were observed in leukocyte transendothelial transplantation. Conclusion: the gene expression of blastocyst / parietal trophoblast cells is revealed from the spatial dimension, which indicates that the blastocyst pole / wall trophoblast ectoderm is coordinated with each other, and the hatching and implantation process of blastocyst are carefully regulated. Further data analysis is expected to find endogenous specific molecules regulating embryo implantation.
【作者單位】： 北京積水潭醫(yī)院婦產(chǎn)科;
【基金】：國家自然科學(xué)基金(81070493)資助~~
【分類號】：R714.8

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