本文選題：囊胚　切入點(diǎn)：極滋養(yǎng)層細(xì)胞　出處：《北京大學(xué)學(xué)報(bào)(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(xì)胞基因的空間表達(dá)。方法:輔助生殖來源人受精后第6天Gardner評(píng)分5AA的囊胚,顯微鏡下微吸管機(jī)械分離滋養(yǎng)外胚層細(xì)胞,對(duì)囊胚滋養(yǎng)外胚層極滋養(yǎng)層細(xì)胞和壁滋養(yǎng)層細(xì)胞采用單細(xì)胞測序檢測,選取經(jīng)t檢驗(yàn)計(jì)算所得P0.05且基因表達(dá)差異≥2倍的差異表達(dá)基因,采用生物信息學(xué)軟件對(duì)篩選的差異基因進(jìn)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(duì)差異表達(dá)基因進(jìn)行注釋和生物功能富集,并用全基因組對(duì)差異基因功能進(jìn)一步注釋。結(jié)果:對(duì)2枚受精后第6天的5AA囊胚滋養(yǎng)外胚層8個(gè)極滋養(yǎng)層細(xì)胞和8個(gè)壁滋養(yǎng)層細(xì)胞測序,發(fā)現(xiàn)極滋養(yǎng)層細(xì)胞306個(gè)基因上調(diào),壁滋養(yǎng)層細(xì)胞75個(gè)基因上調(diào)。無監(jiān)督聚類分析結(jié)果顯示外胚層細(xì)胞分成極滋養(yǎng)層細(xì)胞和壁滋養(yǎng)層細(xì)胞兩群,屬于兩種不同類型和生物功能的細(xì)胞群。差異基因功能注釋顯示,極滋養(yǎng)層細(xì)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(yùn)、氧化應(yīng)激、離子轉(zhuǎn)運(yùn)、蛋白合成及運(yùn)輸、細(xì)胞周期調(diào)節(jié)、肌動(dòng)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(hào)通路、雌雄激素代謝等信號(hào)通路;壁滋養(yǎng)層細(xì)胞上調(diào)基因GO生物學(xué)功能主要是蛋白分解代謝、細(xì)胞周期停滯、細(xì)胞凋亡、激活絲裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信號(hào)通路、碳水化合物運(yùn)輸、突觸負(fù)調(diào)節(jié)、細(xì)胞生長、鈣通道激活、正向B細(xì)胞分化、T細(xì)胞凋亡等,主要參與B細(xì)胞受體、T細(xì)胞受體、白細(xì)胞跨內(nèi)皮移植、血管內(nèi)皮生長因子(vascular endothelial growth factor,VEGF)表達(dá)、間隙連接、促性腺激素釋放激素(gonadotropin-releasing hormone,GnRH)分泌、細(xì)胞凋亡等信號(hào)通路。結(jié)論:從空間維度揭示囊胚極/壁滋養(yǎng)層細(xì)胞的基因表達(dá),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(xì)調(diào)節(jié)囊胚孵化和胚胎植入過程,進(jìn)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(hào)】：R714.8

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