新基因mgt-16对间充质干细胞增殖、迁移的影响及机制研究

项目名称： 新基因mgt-16对间充质干细胞增殖、迁移的影响及机制研究

项目编号： No.31201035

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 王明科

作者单位： 中国人民解放军海军医学研究所

项目金额： 23万元

中文摘要： 间充质干细胞（Mesenchymal stem cells，MSCs）是创伤修复中细胞治疗的最佳候选种子细胞，但其增殖、迁移分子机制有待阐明。课题组前期利用基因捕获从MSCs细胞株10T1/2发现一个新基因mgt-16（已提交GenBank，ID号：GU266552)，已研究其细胞分布、与平滑肌分化的关系，并提示可能与细胞增殖、迁移相关。本课题拟建立过表达或沉默mgt-16的10T1/2细胞模型，用CCK-8、细胞划痕、Transwell、裸鼠皮肤切割伤模型等体内外实验进一步确定mgt-16对细胞增殖、迁移的影响；Tunel及流式检测mgt-16对细胞周期、凋亡的影响；RT-PCR、免疫组化和/或Western blot检测增殖及迁移相关基因的表达，并用信号通路阻滞剂/激动剂等手段，揭示mgt-16参与调控MSCs增殖、迁移的分子机制，为MSCs应用于创伤修复提供理论基础和可能的治疗策略。

中文关键词： 新基因；间充质干细胞；增殖；迁移；分化

英文摘要： Mesenchymal stem cells（MSCs）represent the most promising candicate cell source for cell replacement therapy, which is one of the feasible strategies in the management of wound healing. However, the molecular mechanisms need to be elucidated. This study is based on a novel gene, mgt-16, cloned and characterizated in murine mesenchymal stem cell line 10T1/2 by gene trap screening, which has been submitted to GenBank (GenBank accession no. GU266552).Our primary results showed its cytoplasmic distribution, its roles in smooth muscle cell differentition, and it may be involed in the process of cell proliferation and migration. In this study, we will construct mgt-16 over-expressed and low-expressed 10T1/2 cell models. The influence of mgt-16 on the cell proliferation will be determined using WST-8 dye (Cell Counting Kit-8), cell migration by scratch wound on monolayer cells, Transwell assay, and skin excision wound model in athymic mouse in vitro and in vivo. The influence of mgt-16 on the cell cycle and apoptosis will be observed by Tunel labeling assay and flow cytometry, and cell proliferation or migration related genes expression will be detected by RT-PCR, immunocytochemistry, and western blot in mgt-16 over-expressed or low-expressed 10T1/2 cells. Finally, the molecular mechenisms of how mgt-16 regulats mesench

英文关键词： Novel gene；Mesenchymal stem cells；Cell proliferation；Cell migration；Cell differentiation