我们前期研究发现，胶质瘤高表达CT23与P53及患者预后有关；鉴于CT23启动子含P53结合位点及CT23与TGFβ享有共同靶标miR326，推测P53上调CT23后通过miR326调控TGFβ。故拟开展以下研究:一p53对CT23转录调控：p53-/-细胞转染p53质粒，观察p53对CT23表达影响；用报告基因实验分析p53影响CT23启动子活性；通过EMSA和ChIP-PCR确认p53与CT23启动子结合。二CT23/miR326/TGFβ调控关系及功能：检测CT23/miR326/TGFβ表达并分析相关性；用3’UTR报告基因实验验证miR326靶向结合CT23/TGFβ;借助Dicer酶下调细胞验证CT23对TGFβ调控的miRNA依赖性；构建小鼠移植瘤，从肿瘤细胞、微环境和免疫细胞等方面探讨p53上调CT23诱导TGFβ所致的胶质瘤免疫逃逸机制，为肿瘤靶向药物的靶点选择提供依据。

Our previous study have demonstrated that highly expressed CT23 is related to P53 and patients’ poor prognosis in glioma. As CT23 promoter contains binding sites of P53 ,and CT23 and TGF-β share a common target with miR-326, we infer that up-regulation of CT23 by P53 induces immunologic escape via TGF-β in glioma. Therefore, following study will be performed. Firstly, it is to explore the role of P53 in up-regulation of CT23, which include the transfection of P53 to cells with p53-/- for understanding the influence of P53 on CT23 expression, confirmation of influence of P53 on promoter activity of CT23 with reporter gene assay, and binding test of P53 and CT23 by using EMSA and ChIP-PCR. Secondly, it is to explore the relation and function of CT23/miR326/TGF-β. The expression and relation of CT23/miR326/TGF-β will be tested. The target of miR326 to CT23/TGF-β will be confirmed by using 3’UTR reporter gene assay. By using cells with decrease of Dice enzyme miRNA dependent is confirmed when CT23 regulated TGF-β. Finally, with building up mouse model of transplanted tumor we will explore the mechanism of up-regulation of CT23 by P53 inducing immunologic escape of glioma via TGF-β, which will provide data for selection of tumor targeting treatment.