Senescence bypass in epithelial neoplastic progression

上皮肿瘤进展中的衰老旁路

基本信息

批准号：
7460590
负责人：
JAMES RHEINWALD
金额：
$ 39.35万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-09-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7460590
关键词：
Acute Basement membrane Bypass CDKN2A gene Cell Cycle Inhibition Cell Nucleus Cells Characteristics Chronic Clinical Closure Cultured Cells Diagnosis Dominant-Negative Mutation Drug Combinations Elements Epidermal Growth Factor Receptor Epidermis Epithelial Experimental Models Growth Healed Hemidesmosomes Human Indium Integrins Invasive Lesion Longevity MAP3K7 gene Membrane Proteins Methods Microscopic Modeling Molecular Neoplasms Neoplastic Cell Transformation Nuclear Translocation Oral Oral mucous membrane structure Organ Culture Techniques Pathologic Pathway interactions Pharmaceutical Preparations Phosphorylation Principal Investigator Proteins Rate Retroviral Vector Signal Pathway Skin Small Interfering RNA Specimen Stimulus Stratified Squamous Epithelium Surface Testing Time Tissues Transforming Growth Factor beta Up-Regulation Western Blotting Wound Healing Xenograft Model autocrine base cell motility cellular engineering healing improved in vivo inhibitor/antagonist keratinocyte knock-down laminin-5 neoplastic cell outcome forecast prevent programs receptor research study response senescence surface coating therapeutic target tool tumor progression wound

项目摘要

DESCRIPTION (provided by applicant): Our long-term objective is to characterize the mechanisms of and normal and pathologic settings of growth arrest signal pathways in stratified squamous epithelia, especially the epidermis and oral mucosa. This project seeks to extend our earlier findings about p16INK4A expression in the limited replicative lifespan of human keratinocytes grown in culture and the settings in which this protein becomes expressed in epithelial tissues in vivo. These studies have revealed that p16 and the precursor form of the basement membrane protein Laminin 5 (Lam5pre) are coexpressed in culture by senescing keratinocytes and in vivo by keratinocytes in late dysplastic/early invasive regions of oral mucosa and epidermis and at the migrating fronts of normal healing skin wounds, p16 and Lam5pre are also coexpressed in culture by early passage keratinocytes at the edges of wounds made in confluent cultures and by keratinocytes plated on surfaces coated with the gamma2 precursor form of Laminin 5, associated with growth arrest and directed hypermotility. We have termed this the "keratinocyte motility/arrest (KMA)" response. The arrest and motility features of KMA are uncoupled in p16- and p53-deficient keratinocytes and in SCC cells: such cells become hypermotile when plated on Lam5pre, but are not growth-arrested. KMA is also induced in normal keratinocytes growing on control surfaces by treatment with TGFbeta. Very interestingly, KMA induction KMA by Lam5pre is dependent upon activity of the TGFbeta receptor. These results provide the rationale and experimental tools for investigating the molecular mechanisms of the KMA response, aimed at understanding how KMA is activated in wound repair and how its growth arrest component is evaded during neoplastic progression. We propose to use cell culture, expression and siRNA retroviral vectors, organotypic culture and xenograft models, and microscopic, immunocytologic, immunohistologic, and Western blot methods to elucidate the proteins and signal pathways that trigger the KMA hypermotility/growth arrest response, which appears to be a central element in epithelial wound healing and neoplastic progression.

描述（由申请人提供）：我们的长期目标是表征分层鳞状上皮（尤其是表皮和口服粘膜）中生长停滞信号途径的正常和病理环境的机制。该项目旨在扩展我们对培养物在培养物中生长的人角质形成细胞的有限复制寿命和该蛋白在体内上皮组织中表达的环境中对P16INK4A表达的早期发现。这些研究表明，p16和地下膜蛋白层层粘连蛋白5（LAM5PRE）的前体形式通过角质上的角质细胞和体内的培养物和体内的培养物在培养物中得到共表达，并在晚期发育不良/早期的口服mucosa and epedermiss和Epedermiss和Epedrated Fertrated的培养物中共表达。正常的愈合皮肤伤口，p16和LAM5PRE在培养物中也通过早期通过角质形成细胞在融合培养物中的伤口的边缘以及与粘膜蛋白5覆盖的表面上的角质形成细胞的边缘共表达，与粘蛋白5的表面上，与长粘蛋白蛋白5，与生长滞留和置换性相关。我们将其称为“角质形成细胞运动/逮捕（KMA）”的反应。 KMA的停滞和运动特征在P16和p53缺陷的角质形成细胞和SCC细胞中没有耦合：当将LAM5PRE铺路时，此类细胞会变得高度激动，但没有生长。通过用TGFBETA处理，在对照表面生长的正常角质形成细胞中也诱导KMA。有趣的是，LAM5PRE的KMA诱导KMA取决于TGFBETA受体的活性。这些结果提供了研究KMA反应的分子机制的理由和实验工具，旨在了解KMA在伤口修复中的激活以及在肿瘤进展过程中如何避免其生长阻滞成分。我们建议使用细胞培养，表达和siRNA逆转录病毒载体，器官培养和异种移植模型，以及微观，免疫细胞学学，免疫组织学和蛋白质印迹方法来阐明蛋白质和信号途径，这些蛋白质和信号途径触发KMA高温/生长动作响应，这些途径似乎是在似乎是KMA高温/生长响应。成为上皮伤口愈合和肿瘤进展的中心元素。