Characterisation of Epithelial-Mesenchymal Transition (EMT) in Oral Keratinocytes

导语：口腔中大量寄生着各种各样的微生物群。复杂的细菌生物膜在牙齿的硬组织(牙齿)上形成，特别是在牙周缝隙的根面上，它们与牙缝上皮密切相关。这种上皮显示出与其他身体部位相同的特性，并在保护潜在宿主组织免受细菌入侵方面发挥关键作用。事实上，这种上皮细胞可以协调局部的炎症反应，并释放有助于维持其完整性的防御分子。发生在口腔上皮细胞中的一个潜在的新过程是上皮-间充质转化(EMT)，它导致上皮细胞向间充质表型的转变。虽然EMT在组织形成和修复中是必不可少的，但它在疾病期间的失调会导致长期和长期的细胞因子释放，使细菌组织入侵并导致慢性炎症损害。低水平：主要口腔细菌通过EMT破坏上皮屏障，导致溃疡和无法溶解的慢性炎症损害。该项目将涉及使用永生和原代人类细胞开发EMT模型系统(2D和3D)。随后将使用分子和细胞分析来评估EMT的调节，包括基因和蛋白质表达分析、细胞形态和组织学特征、上皮屏障和细菌入侵研究，这些研究涉及相关的相互作用的免疫系统细胞，例如中性粒细胞。EMT细胞内信号通路(例如，核因子-kB、SMAD)也将在使用药物调节剂的模型系统中进行剖析。主要参考文献：1.米尔沃德。Chapple ILC先生，Wright HJ，Millard JL，Matthews JB，Cooper PR。牙周病原菌对口腔上皮细胞中核因子-kB的激活及基因表达的影响CLIN Exp免疫。2007年5月；148(2)：307-324 2.Prime SS，Nixon SV，Crane IJ.口腔鳞状细胞癌在细胞培养中的行为。J Pathol 1990；160(3)：259-269 3.Khan E，Shelton RM，Cooper PR，Hamburger J，Landini G.H400和原代大鼠角质形成细胞器官型培养的结构特征.《生物医学杂志》2012年6月26日。阿卜杜勒卡里姆；米尔沃德；库珀；谢尔顿；兰迪尼。牙周病原体在体外促进口腔鳞癌细胞的EMT，接受发表在《细胞黏附与迁移2017》上。

INTRODUCTION: The oral cavity is heavily colonised by a diverse microflora. Complex bacterial biofilms develop on the dental hard tissues (teeth) and in particular on the root surface in the gingival crevice where they become intimately associated with the crevicular epithelium. This epithelium, exhibits properties in common with other bodily sites and plays a key role in protecting the underlying host tissues from bacterial invasion. Indeed this epithelium can orchestrate the local inflammatory response and releases defence molecules which help maintain its integrity. A potential novel process that occurs in the oral epithelia is that of epithelial-mesenchymal transition (EMT) which results in the conversion of epithelial cells to a mesenchymal phenotype. While EMT is essential in tissue formation and repair its dysregulation during disease can result in the development of prolonged and chronic cytokine release perpetuating bacterial tissue invasion and resulting in a chronic inflammatory lesion.HYPOTHESIS: Key oral bacteria disrupt the epithelial barrier via EMT resulting in ulceration and a non-resolving chronic inflammatory lesion.The project will involve the development of an EMT model system (2D & 3D) using immortal and primary human cells. EMT regulation will subsequently be assessed using molecular and cellular assays including gene & protein expression analysis, cell morphological & histological characterisation, epithelial barrier and bacterial invasion studies which involve relevant interacting immune system cells, e.g. neutrophils. EMT intracellular signalling pathways (e.g. NF-kB, SMAD) will also be dissected in model systems using pharmaceutical modulators. KEY REFERENCES: 1. Milward. MR, Chapple ILC, Wright HJ, Millard JL, Matthews JB, Cooper PR. Differential Activation of NF-kB and gene expression in oral epithelial cells by periodontal pathogens. Clin Exp Immunol. 2007 May;148 (2): 307-324 2. Prime SS, Nixon SV, Crane IJ. The behaviour of human oral squamous cell carcinoma in cell culture. J Pathol 1990; 160 (3): 259-269 3. Khan E, Shelton RM, Cooper PR, Hamburger J, Landini G. Architectural characterization of organotypic cultures of H400 and primary rat keratinocytes. J Biomed Mater Res A. 2012 Jun 26.4. Abdulkareem; Milward; Cooper; Shelton; Landini. Periodontal pathogens promote EMT in oral squamous carcinoma cells in vitro, accepted for publication in Cell Adhesion & Migration 2017.