细胞周期调控蛋白50A对卵巢癌干细胞的识别作用及机制研究

Epithelial ovarian cancer (EOC) is the leading cause of gynecological cancer mortality. The main reason is the tumor recurrence, metastasis, and chemo-resistance, which is due to the heterogeneity of tumor cells. The research of cancer stem cells may become the key to understand the biology characteristic of ovarian cancer, and change the clinical diagnosis and treatment. Therefore, screening of the specific surface marker of ovarian cancer stem cells is the premise to develop the targeted therapy for ovarian cancer. In the early stage of our work, on the basis of the sorting, enrichment and the culture of side population cells, we used quantitative proteomic techniques and mass spectrometry to find that the expression of CDC50A is higher in side population cells. The further research showed that CDC50A positive cells have the ability of differentiation, sphere formation, self-renewal, and tumorigenesis. Therefore, this project will aim to confirm whether CDC50A can be used as a specific marker of ovarian cancer stem cells and identify whether the CDC50A positive cells in primary ovarian cancer tissues have properties of cancer stem cells. Besides, we will analyze the relationship between the expression of CDC50A protein and the clinical outcomes of EOC patients and further find the signaling pathway of CDC50A through the mRNA sequencing.

卵巢上皮癌女性生殖器官首位致死性疾病，肿瘤细胞的异质性以及由此发生的不断复发、转移、以及对化疗耐药是主要原因。有关肿瘤干细胞（CSC）研究可能成为认识卵巢癌生物学特性形成的关键，并能改变临床诊断和治疗方法。寻找卵巢癌干细胞特异性表面标记物是开辟卵巢癌靶向治疗新靶点的前提。本课题组在对多个卵巢癌细胞系SP细胞分选、富集以及体外培养技术基础上，采用微量蛋白质组学技术和比较蛋白质组分析，发现CDC50A蛋白SP细胞中高表达，进一步通过体外实验证实CDC50A+细胞具有体外分化能力，sphere形成能力，自我更新能力，体内实验证实具有更强的致瘤能力。本课题旨在深入探讨CDC50A作为卵巢癌干细胞特异性标记物的可能性，原代卵巢癌组织中CDC50A+细胞是否具有肿瘤干细胞特性，结合临床资料，分析CDC50A蛋白的表达与卵巢癌患者临床特征及复发的相关性，进一步通过mRNA测序找到其发挥作用的信号通路。

卵巢上皮癌是女性生殖器官首位致死性疾病，肿瘤复发、耐药是临床治疗的主要障碍。前期研究探索证实卵巢癌细胞系CDC50A+细胞具有干细胞相关功能，在此基础之上，本研究成功建立人卵巢癌原代细胞sphere培养体系，能够稳定培养并传代。并在细胞学体外实验、动物实验证实卵巢癌原代CDC50A+细胞具有较强的增殖能、分化能力、自我更新能力、小鼠成瘤能力。此外，还与铂类化疗药物耐药密切相关。通过mRNAs测序，比较CDC50A+和CDC50A-细胞差异表达基因，发现ATP11A/B可能是卵巢癌细胞内CDC50A发挥干细胞相关生物学功能的通路蛋白，并运用分子生物学技术证实ATP11/B参与调节CDC50A+细胞sphere形成能力及对铂类化疗药物的敏感性。