Annexin A1通过调控TGF-β 受体内吞促进人卵巢癌细胞增殖的机制研究

The prognosis of ovarian cancer is the worst in gynecologic malignant tumor. Since conventional treatments are difficult to significantly improve the long-term survival of the patients, the finding of new molecular targets will provide a new way for the treatment of ovarian cancer. In the existing experiment, we produced polyclonal antibodies against SKOV3 cells, which inhibited tumor growth in vitro and in vivo. We used immunoproteomic approach to reveal Annexin A1 (AnxA1) as immunogenic antigens of ovarian cancer. Decreased AnxA1 expression inhibited the activation of TGF-β signaling in ovarian cancer cells and inhibited cell proliferation and invasion. The endocytosis of TGF-β receptor is an important way to regulate TGF-β signaling, and AnxA1 participates in endocytosis of membrane surface proteins. Our project will use co-immunoprecipitation, immunofluorescence, computational analysis experiments, combining with tumor xenograft models and clinical analysis, to study the molecule mechanism of AnxA1 on TGF-β receptor endocytosis, and the increased proliferation caused by TGF-β signaling activation. Our study contributes to bridge the gap between basic research and clinical practice, which provides new ideas for the molecular treatment of ovarian cancer.

卵巢癌在妇科恶性肿瘤中预后最差，常规治疗方式难以明显提高患者的远期生存率，筛选新的分子靶点将为治疗卵巢癌提供新途径。前期实验中，申请者制备的抗人卵巢癌SKOV3细胞多克隆抗体能够在体外和体内实验中抑制肿瘤生长。运用免疫蛋白质组学方法对多克隆抗体识别的卵巢癌抗原进行筛选，确定膜联蛋白A1（Annexin A1，AnxA1）是与卵巢癌密切相关的肿瘤抗原，干扰AnxA1表达抑制卵巢癌细胞TGF-β信号通路，同时抑制细胞增殖与侵袭。鉴于TGF-β受体内吞是调控TGF-β信号水平的重要方式，而AnxA1的经典功能是参与多种膜表面蛋白质内吞，本课题将利用免疫共沉淀、免疫荧光、计算机模拟分析等实验，结合裸鼠移植瘤模型和临床病例分析，研究AnxA1促进TGF-β受体内吞的分子机制，以及TGF-β信号通路激活后对卵巢癌细胞增殖的影响。我们的研究有助于践行转化医学，为卵巢癌分子靶向治疗提供新的思路。

卵巢癌在妇科恶性肿瘤中预后最差，筛选新的分子靶点将为治疗卵巢癌提供新途径。前期实验运用免疫蛋白质组学方法对多克隆抗体识别的卵巢癌抗原进行筛选，确定膜联蛋白A1（Annexin A1，AnxA1）是与卵巢癌密切相关的肿瘤抗原，目前已完成慢病毒载体LV-AnxA1（AnxA1 过表达载体）和LV-shAnxA1（AnxA1 RNA 干扰载体）的构建；慢病毒感染SKOV3 细胞及稳定转染细胞的筛选；体外实验研究过表达AnxA1 蛋白促进TGF-β信号激活及相关细胞学效应；完成动物体内实验研究；研究AnxA1 与卵巢癌患者病理特征关系；AnxA1与卵巢癌患者生存及预后关系。目前AnxA1 促进TGF-β受体内吞分子机制的研究还在继续进行中。AnxA1与卵巢癌关系的进一步研究将为后续小分子抑制剂，单克隆抗体，抗体药物偶联物(ADC) 的研发奠定重要的理论基础。