大鼠气管干细胞增殖分化过程中ABCG2的表达及Wnt信号机制的研究

发布时间：2018-05-08 00:30

本文选题：ABCG2 + 气管干细胞　；参考：《中国医科大学》2006年博士论文

【摘要】：前言干细胞是指具有无限或较长期的自我更新能力,并能产生至少一种高度分化子代细胞的细胞。按照组织发生学来源,可将干细胞分为胚胎干细胞和成体干细胞。近年来对成体干细胞的研究越来越受到重视,目前已发现的成体干细胞有造血干细胞、骨髓间充质干细胞、神经干细胞、肌干细胞、角膜干细胞、胰腺干细胞、皮肤干细胞、肝脏干细胞等,并已不同程度地用于临床治疗。但迄今为止国际上对气管干细胞的研究还很不完善。先前本研究组已成功地在体内外建立了氟脲嘧啶介导的大鼠气管损伤修复模型,并对气管干细胞进行了初步的定位和性状解析。干细胞具有排出荧光染料Hoechst33342的特性。近年来的研究表明,干细胞的这一特性(即SP表型)与ATP结合盒(ABC)转运蛋白超家族成员之一的ABCG2(又名乳癌耐药蛋白1,Bcrp1)的表达有关。现已证明在多个物种的多种组织来源的SP细胞中检测到ABCG2的高表达,如骨髓、肝、肌组织、乳腺上皮及ES细胞。ABCG2与SP表型的密切关系,使得ABCG2成为多潜能干细胞的共同标志,对干细胞的鉴定具有重要意义。为更深入地了解气管干细胞的特性,本研究在5-FU介导的气管上皮损伤修复模型的基础上,应用间接免疫荧光法和Wetern blot动态观测了修复过程中各时间点气管上皮ABCG2蛋白的表达,借以原位观察了气管干细胞在整个修复过程中的动态分布。近年来的研究表明Wnt信号及其成员对胚胎及成体的原始细胞的增殖和分化具有重要的调节作用。为探讨气管干细胞的增殖分化与Wnt/β-catenin信号途径的关系,阐明干细胞增殖分化的分子机制,本研究应用RT-PCR及间接免疫荧光的方法动态观测了修复过程中Wnt信号相关成分Wnt1、β-catenin、cyclinD1 mRNA的变化以及β-catenin蛋白的细胞内分布,初步探讨了Wnt/β-catenin信号途径参与调控气管干细胞增殖
[Abstract]:Preface Stem cells are cells that have unlimited or long-term self-renewal and produce at least one highly differentiated progeny. Stem cells can be divided into embryonic stem cells and adult stem cells according to the origin of histogenesis. In recent years, more and more attention has been paid to the research of adult stem cells. At present, the adult stem cells have been found to be hematopoietic stem cells, bone marrow mesenchymal stem cells, neural stem cells, muscle stem cells, corneal stem cells, pancreatic stem cells, skin stem cells. Liver stem cells and so on, and have been used in clinical treatment to varying degrees. However, the international research on tracheal stem cells is not perfect. The model of rat trachea injury induced by fluorouracil in vitro and in vivo has been successfully established, and the primary localization and character analysis of tracheal stem cells have been carried out. Stem cells are characterized by the emission of fluorescent dye Hoechst33342. Recent studies have shown that this characteristic of stem cells (SP phenotype) is related to the expression of ABCG2 (also known as Bcrp1), a member of the ATP binding cassette transporter superfamily. It has been proved that high expression of ABCG2 has been detected in SP cells from many species, such as bone marrow, liver, muscle, mammary epithelium and es cells. ABCG2 is closely related to SP phenotype, which makes ABCG2 a common marker of multipotential stem cells. It is of great significance for the identification of stem cells. In order to better understand the characteristics of tracheal stem cells, the expression of ABCG2 protein in tracheal epithelium at different time points was dynamically observed by indirect immunofluorescence and Wetern blot on the basis of 5-FU mediated repair model of tracheal epithelial injury. The dynamic distribution of tracheal stem cells during the whole repair process was observed in situ. Recent studies have shown that Wnt signaling and its members play an important role in regulating the proliferation and differentiation of embryonic and adult primordial cells. To investigate the relationship between proliferation and differentiation of tracheal stem cells and Wnt/ 尾 -catenin signaling pathway, and to elucidate the molecular mechanism of stem cell proliferation and differentiation. In this study, RT-PCR and indirect immunofluorescence were used to dynamically observe the changes of Wnt signal related components Wnt1, 尾 -catenin D1 mRNA and the intracellular distribution of 尾 -catenin protein, and to explore the role of Wnt/ 尾 -catenin signaling pathway in regulating the proliferation of tracheal stem cells.
【学位授予单位】：中国医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R329

【参考文献】