癌旁成纤维细胞的募集和分化是其促进肿瘤发生发展必不可少的两个重要生物学行为，申请者领导的课题组前期研究发现：成纤维细胞在迁移的同时其分化将受到抑制，并首次提出了癌旁成纤维细胞存在初始分化-募集-完全分化的"两阶段分化"时相特征且受到"TGF-β时相分子开关"调控的假说。本项目通过建立口腔癌体外三维培养模型和SCID小鼠模型，联合应用实时图像采集、激光共聚焦、流式微球分析、RNA干扰、IP-Western等生物学技术研究 （1）口腔癌旁成纤维细胞募集－分化的时相特征;（2）TGF-β刺激口腔癌旁成纤维细胞，募集和分化的信号调控通路及其分子机制；（3）癌旁成纤维细胞募集-分化信号调控通路的交互对话及"时相分子开关"；为验证"TGFβ-成纤维细胞两阶段分化"假说，阐明口腔上皮－间质交互作用的分子机制，寻找阻断口腔癌的新策略和新靶点提供实验依据和理论基础，具有重要的理论意义和广阔的应用前景。

The recruitment and the differentiation of the surrounding fibroblasts in oral cancer, the two important biological behaviors, show essential for the tumor development. The preliminary study of the group leaded by the applicant found that the differentiation of the fibroblasts was inhibited when migrating. The hypotheses are first proposed that the surrounding fibroblasts appear the two-stage differentiation pattern and time phase of initial differentiation, recruitment and complete differentitiation, which might be regulated by the time phase molecular swich of TGF-β. The techniques adopted by the project include three-dimensional culture in vitro model and SCID mouse model of oral cancer, real-time image acquisition, flow cytometric bead analysis, RNA interference, immunoprecipitation and western blotting. The research involves the pattern and time phase of the recruitment and the differentiation of the surrounding fibroblasts in oral cancer, the regulatory pathway and its mechanism of the recruitment and the differentiation by TGF-β, the cross talk between the regulatory pathways of the recruitment and the differentiation and the time phase molecular swich. The project can test the hypothesis of the two-stage differentiation of the fibroblasts by TGF-β, can clarify the molecular mechanism of oral epithelial-stromal interaction, can provide experimental basis and theoretical basis for seeking the new strategies and new targets to block the development of oral cancer and so it possesses important theoretical significance and potential applications.