整合素β1在人表皮细胞分化中作用的研究
发布时间：2018-12-12 03:25
【摘要】： 一、研究背景 各种损伤特别是大面积烧伤导致皮肤缺损的治疗,是尽快封闭创面,由于自体皮源的不足,常导致治疗的失败。组织工程化皮肤研究一直是当前研究的热点,目前尚无特别有效的皮肤替代物应用于临床。而表皮干细胞(keratinocyte stem cell, KSC)具有强大的增殖能力,又可向各个阶段表皮细胞分化以维持皮肤新陈代谢,因此KSC作为皮肤重建的“种子细胞”成为研究的热点。如何在体外培养获得足够数量的KSC用于研究及移植,如何在体内外诱导KSC定向分化,对于组织工程皮肤具有重要意义。解决这些问题的关键在于研究KSC增殖与分化的调控机制。整合素β1是KSC未分化的分子标记之一,Jones等发现高表达整合素β1的表皮细胞具有更高的克隆形成能力,传代次数显著多于低表达的细胞[1]。。本室前期通过构建整合素β1小RNA干扰载体,转染KSC,发现下调表达整合素β1蛋白,实验组克隆形成率比对照组降低,促进KSC的分化[2]。提示整合素β1参与KSC的增殖分化的调控。目前虽然对整合素β1参与KSC增殖与分化的调控研究比较多,但是整合素β1的表达调控的机制尚不清楚。 体外实验研究中,KSC通常从不同的个体分离和纯化,存在个体差异和不稳定性,因此寻找一种稳定细胞模型对KSC进行分化与增殖调控机制的研究具有重要意义。HaCaT是来源于正常成人上皮细胞的表皮细胞株,具有无限的增殖能力。研究表明,将HaCaT移植到裸鼠上,HaCaT可以形成分化上皮组织,表达特异性分化蛋白标志,因此HaCaT可以为研究人表皮细胞分化的调控提供一个非常理想的模型[3]。研究拟应用整合素β1启动子荧光素酶报告基因系统,以HaCaT细胞为模型,探讨表皮细胞中整合素β1对增殖分化的影响,以及整合素β1的表达调节机制,为进一步研究表皮干细胞的增殖分化机制奠定基础。 二、研究目的 1、下调人表皮细胞株HaCaT整合素β1,观察其增殖的情况。 2、研究整合素β1启动子在HaCaT中的活性,为研究干细胞分化与增殖调控机制提供理论基础。
[Abstract]:Background: the treatment of skin defect caused by various kinds of injuries, especially large area burn, is to close the wound as soon as possible, because of the insufficiency of autogenous skin source, which often leads to the failure of treatment. Tissue engineered skin research has been the focus of current research, and there is no especially effective skin substitute for clinical application. The epidermal stem cell (keratinocyte stem cell, KSC) has strong proliferative ability and can differentiate into epidermal cells to maintain skin metabolism. Therefore, KSC as a "seed cell" for skin reconstruction has become a hot research topic. How to obtain enough KSC in vitro for research and transplantation, and how to induce directional differentiation of KSC in vitro and in vivo are of great significance for tissue engineering skin. The key to solve these problems is to study the regulation mechanism of KSC proliferation and differentiation. Integrin 尾 1 is one of the undifferentiated molecular markers of KSC. Jones et al found that the epidermal cells with high integrin 尾 1 expression had higher clone forming ability, and the number of passages was significantly higher than that of low expression cells [1]. In the early stage of this study, integrin 尾 1 small RNA interference vector was constructed, and down-regulated expression of integrin 尾 1 protein was found in KSC, transfection. The clone formation rate of the experimental group was lower than that of the control group, and the differentiation of KSC was promoted [2]. These results suggest that integrin 尾 1 is involved in the regulation of KSC proliferation and differentiation. Although there are many studies on integrin 尾 1 involved in KSC proliferation and differentiation, the mechanism of integrin 尾 1 expression regulation is not clear. In vitro, KSC is usually isolated and purified from different individuals. Therefore, it is of great significance to study the mechanism of differentiation and proliferation of KSC in a stable cell model. HaCaT is an epidermal cell line derived from normal adult epithelial cells and has unlimited proliferative ability. The results showed that HaCaT could form differentiated epithelial tissue and express specific differentiation protein markers by transplanting HaCaT to nude mice. Therefore, HaCaT could provide a very ideal model for studying the regulation of differentiation of human epidermal cells [3]. To study the effect of integrin 尾 1 on proliferation and differentiation and the regulation mechanism of integrin 尾 1 expression in epidermal cells using integrin 尾 1 promoter luciferase reporter gene system as a model of HaCaT cells. It lays a foundation for further study on the mechanism of proliferation and differentiation of epidermal stem cells. Objective 1. To down-regulate HaCaT integrin 尾 1 and observe its proliferation. 2. To study the activity of integrin 尾 1 promoter in HaCaT, and to provide a theoretical basis for studying the mechanism of stem cell differentiation and proliferation.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329
本文编号：2373811
[Abstract]:Background: the treatment of skin defect caused by various kinds of injuries, especially large area burn, is to close the wound as soon as possible, because of the insufficiency of autogenous skin source, which often leads to the failure of treatment. Tissue engineered skin research has been the focus of current research, and there is no especially effective skin substitute for clinical application. The epidermal stem cell (keratinocyte stem cell, KSC) has strong proliferative ability and can differentiate into epidermal cells to maintain skin metabolism. Therefore, KSC as a "seed cell" for skin reconstruction has become a hot research topic. How to obtain enough KSC in vitro for research and transplantation, and how to induce directional differentiation of KSC in vitro and in vivo are of great significance for tissue engineering skin. The key to solve these problems is to study the regulation mechanism of KSC proliferation and differentiation. Integrin 尾 1 is one of the undifferentiated molecular markers of KSC. Jones et al found that the epidermal cells with high integrin 尾 1 expression had higher clone forming ability, and the number of passages was significantly higher than that of low expression cells [1]. In the early stage of this study, integrin 尾 1 small RNA interference vector was constructed, and down-regulated expression of integrin 尾 1 protein was found in KSC, transfection. The clone formation rate of the experimental group was lower than that of the control group, and the differentiation of KSC was promoted [2]. These results suggest that integrin 尾 1 is involved in the regulation of KSC proliferation and differentiation. Although there are many studies on integrin 尾 1 involved in KSC proliferation and differentiation, the mechanism of integrin 尾 1 expression regulation is not clear. In vitro, KSC is usually isolated and purified from different individuals. Therefore, it is of great significance to study the mechanism of differentiation and proliferation of KSC in a stable cell model. HaCaT is an epidermal cell line derived from normal adult epithelial cells and has unlimited proliferative ability. The results showed that HaCaT could form differentiated epithelial tissue and express specific differentiation protein markers by transplanting HaCaT to nude mice. Therefore, HaCaT could provide a very ideal model for studying the regulation of differentiation of human epidermal cells [3]. To study the effect of integrin 尾 1 on proliferation and differentiation and the regulation mechanism of integrin 尾 1 expression in epidermal cells using integrin 尾 1 promoter luciferase reporter gene system as a model of HaCaT cells. It lays a foundation for further study on the mechanism of proliferation and differentiation of epidermal stem cells. Objective 1. To down-regulate HaCaT integrin 尾 1 and observe its proliferation. 2. To study the activity of integrin 尾 1 promoter in HaCaT, and to provide a theoretical basis for studying the mechanism of stem cell differentiation and proliferation.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329
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相关期刊论文 前3条
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