肿瘤微环境对于肿瘤发生发展有着重要的作用，并受到代谢的影响。跨膜糖蛋白的糖基化修饰和糖代谢变化可影响肿瘤微环境。我们发现CD147在黑素瘤原位灶和转移灶中呈不同分布，与其翻译后修饰相关。CD147可从细胞粘附、微环境pH值、细胞因子自分泌等多途径影响细胞微环境。我们拟通过酵母双杂交、IP-质谱等研究方法，鉴定CD147在胞浆中的相互作用蛋白，再通过一系列的分子/细胞生物学后续实验对其胞浆功能进行研究。利用IP-质谱分析临床样本胞浆中CD147大幅度增加后的翻译后修饰变化，构建相应的点突变小鼠黑素瘤模型，验证细胞实验结果。利用原代细胞系、患者来源组织的移植瘤模型（PDX模型）和病理免疫组化等手段结合病人生存率，对细胞和小鼠中的实验结果进行验证。本项目将探讨翻译后修饰（糖基化，泛素化）对于CD147细胞定位改变的分子机制，阐明胞浆CD147的功能及对细胞微环境和黑素瘤侵袭转移的影响。

Tumor microenvironment plays critical role in tumorigenesis,which is controlled by metabolism. The glycosylation of molecules in membrane and cellular glycolysis orchestrate formation of tumor microenvironment. Our preliminary data showed CD147 majorly distributed in cytoplasm in metastatic melanoma tissue comparing to membrane distribution in primary melanoma. We proposed glycosylation alteration of CD147 plays a key role in this phenomena. It was reported that CD147 effects on cell adhesion, pH value of microenvironment, cytokine secretion which affects on tumor microenvironment. We have plan to identify the CD147 interaction proteins in cytoplasm through yeast two hybrid and IP-MS and study the cytoplasmic CD147 function. The glycosylation sites of CD147 in clinical metastatic melanoma tissue would be identified by IP-MS. Mutation of CD147 at glycosylation sites would be generated and be constructed into melanoma animal model to investigate the glycosylation function of CD147 in melanoma. Combination of primary cultured melanoma cell lines and PDX model will be performed to confirm results from melanoma cell lines. The purpose of this application is how glycosylation affects on CD147 cytoplasmic distribution and function, which promotes melanoma progression through tumor microenvironment.