免疫蛋白质组学筛选卵巢癌肿瘤抗原及其功能研究

Ovarian cancer is the leading cause of death among gynecologic malignancies. Despite improvements in surgery and chemotherapy, the overall survival rate of patients has not improved substantially. Molecular targeted therapy offers new avenues for research, and identification of novel molecular marker may provide a potential target for therapeutic intervention. In our study, we produced polyclonal antibodies by immunizing rabbit with human ovarian cancer cell line SKOV3. The polyclonal antibodies inhibited proliferation and induced apoptosis in SKOV3 cells. Meanwhile, administration of polyclonal antibodies reached significant inhibition against tumor xenografts in athymic mice. In the further study, we plan to use immunoproteomic approach to reveal immunogenic antigens of ovarian cancer, which were confirmed by the Protein Bank and clinical data. We choose the target protein, which is closely related with ovarian cancer, and investigate its role in tumorigenesis by using lentiviral vectors expressing target gene or small interfering RNA. Our study may provide a novel therapy target for ovarian cancer in clinical application.

卵巢癌是妇科恶性肿瘤中预后最差的，常规手术及化疗均难以明显提高患者的远期生存率。分子靶向治疗为目前研究热点，筛选新的分子靶点将为治疗卵巢癌提供新途径。申请者利用人卵巢癌SKOV3细胞免疫新西兰大白兔，纯化血清获得多克隆抗体。前期实验发现，制备的多克隆抗体能够与卵巢癌细胞特异性结合，在体外诱导肿瘤细胞凋亡，在裸鼠卵巢癌肿瘤模型中抑制实体瘤生长。我们拟在进一步的研究中，用免疫蛋白质组方法对多克隆抗体识别的卵巢癌相关抗原进行筛选，结合蛋白质功能数据库和临床卵巢癌标本检测，确定与肿瘤密切相关的目标蛋白质。通过慢病毒介导的基因过表达和RNA干扰技术，深入研究目标蛋白质在卵巢癌发生发展中的作用。我们的研究将为卵巢癌的分子靶向治疗提供实验基础和新的思路。

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，分子靶向治疗为目前研究热点，筛选新的分子靶点将为治疗卵巢癌提供新途径。课题研究者利用人卵巢癌SKOV3 细胞免疫新西兰大白兔，纯化血清制备多克隆抗体，检测抗人卵巢癌SKOV3细胞多克隆抗体体外和体内实验中抑瘤活性，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，并通过免疫印迹、免疫荧光、细胞学实验、临床病理标本检测等深入研究目标蛋白质在卵巢癌发生发展中的作用，探索可能的分子机制。我们发现抗人卵巢癌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。AnxA1在临床卵巢癌标本中高表达，干扰AnxA1表达抑制体外卵巢癌细胞TGF-β信号通路，同时抑制细胞增殖与侵袭。我们以多克隆抗体抗肿瘤作用为基础，建立了功能导向的免疫蛋白质组学技术，可以更有效高通量筛选肿瘤抗原，为卵巢癌及其他恶性肿瘤分子靶向治疗提供新的思路。