Wnt信号分子在大鼠气管干细胞增殖分化过程中的表达

发布时间：2018-05-12 11:54

  本文选题：Wnt-1 + β-Catenin　； 参考：《中国医科大学》2006年博士论文

【摘要】：前言 干细胞是具有保持自我复制能力且在适合微环境下具有多分化能力的原始细胞。研究干细胞可以探索人体发生发育之谜,更可望将来人工制造替代患病的组织器官。成体干细胞为最近几年提出的新概念,成体干细胞主要分化成自身组织的细胞,与组织的生长分化修复密切相关。其中,造血、神经、皮肤、肝脏、胰脏、前列腺、胃肠干细胞已相继被证明。 本研究组曾建立了体外观察气管干细胞的模型,并已证明其特点是未分化、对5-FU有耐药性的Go期细胞,能排除Hoechst33342染料,SP性质可通过Bcrp1/ABCG_2基因表达来证明。在气管损伤修复过程中,涉及不同的分子信号级联,控制干细胞的迁移、增生及分化。然而目前国内外尚无此方面的报导。 Wnt蛋白是控制细胞生长、增殖的关键分泌信号分子,可传递细胞间相互调控信息。近年来研究证实,干细胞和细胞外基质的互相作用需要Wnt信号途径的参与。Wnt通过阻止β-catenin的分解,激活转录因子Tcf/Lef的表达,促进干细胞的增殖与分化。 本研究应用蛋白印迹、RT-PCR及免疫组化方法检测了大鼠气管损伤修复中Wnt-1、β-catenin、Tcf-4、c-Myc基因的表达变化情况,探讨Wnt信号途径在气管干细胞增殖分化中的重要作用,希望能对了解诱导气管干细胞增殖分化的分子机制有所帮助。 方法 1 气管损伤模型建立取大鼠气管,剪成2-3mm宽的气管环,分两组。实验组:将气管环置于含5-FU(12.5mg/ml)的Hams F-12液中,5%CO_2,37℃孵育;12小时后弃去上述培养液,换成新鲜的Hams F-12液
[Abstract]:Preface Stem cells are primordial cells that have the ability to maintain self-replication and have the ability to differentiate in a suitable microenvironment. The study of stem cells can explore the mystery of the development of human body, and it is expected to replace the diseased tissues and organs by artificial manufacture in the future. Adult stem cells are a new concept put forward in recent years. Adult stem cells mainly differentiate into cells of their own tissues which are closely related to tissue growth differentiation and repair. Among them, hematopoietic, nerve, skin, liver, pancreas, prostate, gastrointestinal stem cells have been proved. Our team has established an in vitro model of observing tracheal stem cells, and it has been proved that the characteristics of these cells are undifferentiated. The G _ o phase cells resistant to 5-FU can exclude the nature of Hoechst33342 dyestuff SP, which can be proved by the expression of Bcrp1/ABCG_2 gene. In the process of tracheal injury repair, different molecular signal cascades are involved to control the migration, proliferation and differentiation of stem cells. However, there is no such report at home and abroad. Wnt protein is a key secretory signal molecule to control cell growth and proliferation, which can transmit intercellular regulation information. Recent studies have demonstrated that the interaction between stem cells and extracellular matrix requires the involvement of Wnt signaling pathway. Wnt promotes the proliferation and differentiation of stem cells by blocking the decomposition of 尾 -catenin and activating the expression of transcription factor Tcf/Lef. In this study, Western blot RT-PCR and immunohistochemistry were used to detect the expression of Wnt-1, 尾 -cateninine Tcf-4c-Myc gene in trachea injury repair in rats, and to explore the important role of Wnt signaling pathway in the proliferation and differentiation of tracheal stem cells. It may be helpful to understand the molecular mechanism of inducing proliferation and differentiation of tracheal stem cells. Method 1 trachea injury model was established in rats. The trachea was cut into 2-3mm wide trachea rings and divided into two groups. Experimental group: the trachea ring was placed in Hams F-12 solution containing 5-FU 12.5 mg / ml. After incubating at 37 鈩，

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