MECHANISMS OF KERATINOCYTE MOTILITY

描述：(改编自申请人的摘要)-人类 角质形成细胞运动是创伤过程中的关键早期事件 治愈。直接影响人类的生物元素 对角质形成细胞的迁移知之甚少。此外，事实是 角质形成细胞的迁移必须发生在 愈合的伤口决定了实验应该直接设计成 检查伤口环境的要素，并评估它们如何 影响细胞的生物行为。他们的分析方法是专门设计的 从其他生物学角度剖析角质形成细胞的运动功能 细胞增殖等功能。他们最近的工作证明 显然，结缔组织成分或成分的类型 与基底层角质形成细胞并列是诱导的主要影响因素。 迁移和停止迁移。除了细胞外基质 效应，角质形成细胞迁移可受选定生长的影响 因子(如EGF、TGFA和IL-1)，但不受许多其他因素的影响 可溶性因子(如NGF、IL-8、TGFb等)。缺氧也是一个主要原因 影响角质形成细胞迁移的因素。在人类皮肤创伤中 用半透气性闭塞敷料治疗，已经 证明可以促进再上皮化和伤口愈合， 愈合皮肤伤口的微环境以极低的氧气为特征 张力大，pH低。根据这一体内观察，他们的 最近的初步观察表明，角质形成细胞表现出 在低氧条件下加强迁移。在这项提案中，他们希望 探讨人角质形成细胞迁移的细胞机制 结缔组织成分。此外，他们还希望检查这些 常氧和低氧条件下的细胞机制 在微型环境中验证它们的生物学重要性 环境类似于真正的伤口愈合。具体目标是：(一) 检验层粘连蛋白和层粘连蛋白异构体的作用(层粘连蛋白5=外膜蛋白， 对角质形成细胞运动的影响，并确定是否存在低氧 改变角质形成细胞合成和沉积基质黏附 分子，(Ii)确定片状脂膜相关分子的作用 角质形成细胞迁移和迁移中的蛋白质(Ezrin、moesin、Radioxin) 确定低氧对这些基因表达的影响 片状脂膜相关成分，(Iii)检测整合素受体 常氧和低氧条件下的运动性和非运动性角质形成细胞 以及(Iv)研究金属蛋白酶及其受体的作用。 角质形成细胞迁移过程中的各自抑制物和 确定低氧和常氧条件是否会改变它们的表达。

DESCRIPTION: (Adapted from the applicant's abstract) - Human keratinocyte motility is a critical early event in the process of wound healing. The biological elements that directly influence human keratinocyte migration are poorly understood. Moreover, the fact that keratinocyte migration must occur within the microenvironment of the healing wound dictates that experiments be designed that directly examine elements of the wound environment and evaluate how they influence the biological behavior of cells. Their assays are designed to dissect the functions of keratinocyte motility from other biological functions such as cell proliferation. Their recent work demonstrates clearly that the type of connective tissue component or components juxtaposed to the basal keratinocyte is a major influence upon inducing migration and stopping migration. In addition to extracellular matrix effects, keratinocyte migration can be influenced by selected growth factors (such as EGF, TGFA, and IL-1) but not by a number of other soluble factors (e.g., NGF, IL-8, TGFb, etc.). Hypoxia is also a major element in influencing keratinocyte migration. In human skin wounds treated with semi-permeable occlusive dressings that have been demonstrated to promote re-epithelialization and wound healing, the healing skin wound micro-environment is characterized by very low oxygen tension and low pH. In accordance with this in vivo observation, their recent preliminary observations demonstrate that keratinocytes exhibit enhanced migration under hypoxic conditions. In this proposal, they wish to explore the cellular mechanisms of human keratinocyte migration over connective tissue components. Moreover, they wish to examine these cellular mechanisms under normoxic and hypoxic conditions in order to validate their biological importance in the setting of a micro- environment akin to true wound healing. The specific aims are (i) to examine the role of laminin and laminin isoforms (laminin 5 = epiligrin, kalinin, nicein) in keratinocyte motility and determine if hypoxia alters the keratinocyte synthesis and deposition of matrix-adherence molecules, (ii) to determine the role of lamellipodia-associated proteins (ezrin, moesin, radixin) in keratinocyte migration and determine the influence of hypoxia upon the expression of these lamellipodia-associated components, (iii) to examine integrin receptors in motile and non-motile keratinocytes under normoxic and hypoxic conditions, and (iv) to examine the role of metalloproteinases and their respective inhibitors in the process of keratinocyte migration and determine if hypoxic and normoxic conditions alter their expression.