4.1N诱导失巢凋亡抑制上皮性卵巢癌腹腔播散分子机制研究

Epithelial ovarian cancer (EOC) is a highly metastatic disease and the leading cause of death from gynecologic malignancy. The poor prognosis is attributed to its commonly subtle symptoms until widespread intra-abdominal dissemination and metastasis, in which anoikis resistence has been recognised as a key step.Protein 4.1 family members are essential cortical cytoskeleton components underlying the cell membrane and influencing cell adhesion, polarity, migration and response to extracellular signals. We originally found that 4.1N could greatly inhibit peritoneal dissemination of EOC cells by inducing anoikis.Our difference protein mass spectrum results showed that 4.1N might closely involved in the regulation network of integrins.By means of transfection assays, RNA integerence assays,nude mice orthotopic xynograt model, proteomics, bioniformatics and protein-protein interaction assays, our project plan to verify that protein 4.1N induces anoikis by regulating integrin at focal adhesion and hence obviously suppresses the peritoneal implantation of EOCs.This study is trying to integrate a variety of data shown by molecular, cellular, tissue, organ and whole animal body assays, so that we can add some novel insights into the mechanism of anoikis from the perspective of membrance protein localization defect of integrins, and provide a potentially useful molecular prognostic marker and therapeutic target for EOCs.

早期盆腔种植播散是上皮性卵巢癌高致死率主因之一,逃逸失巢凋亡是种植转移的重要步骤。细胞骨架连接蛋白4.1家族是细胞皮质的信号集线器，密切参与多种质膜分子定位和功能调控。申请人原创性发现外源性过表达4.1N蛋白能够诱导卵巢癌细胞发生失巢凋亡并明显抑制卵巢癌腹腔种植转移。差异蛋白质组学分析结果提示，4.1N可能参与调控失巢凋亡核心分子-整合素的信号网络。本立项拟从临床病理实践出发、利用基因重组技术、结合质谱及生物信息学分析、运用活细胞工作站和蛋白-蛋白相互作用技术，对人体肿瘤标本、肿瘤细胞系和动物模型进行整合医学研究。明确4.1N通过调控整合素诱导失巢凋亡、进而抑制上皮性卵巢癌腹腔种植转移的分子机制，为整合素调控网络提供新的节点蛋白，丰富失巢凋亡调控机制，为卵巢癌诊治提供新的靶标。

早期盆腔种植播散是上皮性卵巢癌高致死率主因之一,逃逸失巢凋亡是种植转移的重要步骤。细胞骨架连接蛋白 4.1 家族是细胞皮质的信号集线器，密切参与多种质膜分子定位和功能调控。申请人原创性发现外源性过表达 4.1N 蛋白能够诱导卵巢癌细胞发生失巢凋亡并明显抑制卵巢癌腹腔种植转移。我们发现4.1N的缺失促进上皮性卵巢癌的侵袭、转移和EMT的表达，并促进失巢凋亡的耐受；体内实验验证4.1N缺失促进肿瘤的形成和转移；4.1N通过与14-3-3ζ共定位抑制 14-3-3ζ表达从而抑制上皮性卵巢癌的侵袭、转移、EMT和失巢凋亡；4.1N依赖14-3-3ζ的表型是通过抑制pARP2和cdc42/RAC1通路的激活实现的；4.1N与14-3-3ζ在卵巢癌组织中的表达呈负相关。明确 4.1N 通过调控14-3-3ζ诱导失巢凋亡、进而抑制上皮性卵巢癌腹腔种植转移，丰富失巢凋亡调控机制，为卵巢癌诊治提供新的靶标。