本文選題：Wnt-1 + β-Catenin�。� 參考：《中國醫(yī)科大學(xué)》2006年博士論文

【摘要】：前言 干細(xì)胞是具有保持自我復(fù)制能力且在適合微環(huán)境下具有多分化能力的原始細(xì)胞。研究干細(xì)胞可以探索人體發(fā)生發(fā)育之謎,更可望將來人工制造替代患病的組織器官。成體干細(xì)胞為最近幾年提出的新概念,成體干細(xì)胞主要分化成自身組織的細(xì)胞,與組織的生長分化修復(fù)密切相關(guān)。其中,造血、神經(jīng)、皮膚、肝臟、胰臟、前列腺、胃腸干細(xì)胞已相繼被證明。 本研究組曾建立了體外觀察氣管干細(xì)胞的模型,并已證明其特點是未分化、對5-FU有耐藥性的Go期細(xì)胞,能排除Hoechst33342染料,SP性質(zhì)可通過Bcrp1/ABCG_2基因表達(dá)來證明。在氣管損傷修復(fù)過程中,涉及不同的分子信號級聯(lián),控制干細(xì)胞的遷移、增生及分化。然而目前國內(nèi)外尚無此方面的報導(dǎo)。 Wnt蛋白是控制細(xì)胞生長、增殖的關(guān)鍵分泌信號分子,可傳遞細(xì)胞間相互調(diào)控信息。近年來研究證實,干細(xì)胞和細(xì)胞外基質(zhì)的互相作用需要Wnt信號途徑的參與。Wnt通過阻止β-catenin的分解,激活轉(zhuǎn)錄因子Tcf/Lef的表達(dá),促進(jìn)干細(xì)胞的增殖與分化。 本研究應(yīng)用蛋白印跡、RT-PCR及免疫組化方法檢測了大鼠氣管損傷修復(fù)中Wnt-1、β-catenin、Tcf-4、c-Myc基因的表達(dá)變化情況,探討Wnt信號途徑在氣管干細(xì)胞增殖分化中的重要作用,希望能對了解誘導(dǎo)氣管干細(xì)胞增殖分化的分子機(jī)制有所幫助。 方法 1 氣管損傷模型建立取大鼠氣管,剪成2-3mm寬的氣管環(huán),分兩組。實驗組:將氣管環(huán)置于含5-FU(12.5mg/ml)的Hams F-12液中,5%CO_2,37℃孵育;12小時后棄去上述培養(yǎng)液,換成新鮮的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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