肿瘤靶向多肽受体介导上皮性卵巢癌耐药与转移的作用机制及其靶向阻遏研究

Despite advances in surgical technologies and the development of more effective chemotherapeutics, epithelial ovarian cancer (EOC) remains the leading cause of death in women with gynecologic malignancies. Most of patients with advanced ovarian cancer experience recurrence within five years and eventually die because their cancer becomes resistant to platinum and taxane. Recent advances in our understanding of the molecular biology of cancer and carcinogenesis have led to a variety of targeted agents. The increased characterization of the molecular mechanisms involved in the development and progression of ovarian cancer, and how this has resulted in improved therapeutic strategies with molecular-targeted agents. Our understanding of the molecular pathogenesis and heterogeneity of EOC hold promise for the development of novel therapies against these tumors. Therefore, new treatment strategies are needed to improve outcomes for patients with advanced and recurrent ovarian cancer. The mechanisms underlying the correlation between metastatic potential, coexpression of multiple drug resistance (MDR), and the other cellular functions are unknown and arouse great interest of researchers. In the preliminary unpublished data, we demonstrated TMTP1 specifically bound to highly metastatic ovarian tumor cells and ovarian cancer cisplatin-resistant cell in vitro and in vivo. In addition, it is important to identify the receptor (APP2) that TMTP1 binds at the cell surface and bind to HSP27 in tumor cells, which will help us to better understand the molecular properties of TMTP1. We try to identify the scientific hypothesis that the APP2 gene is a new gene involving the pathogenicity of EOC and explore whether APP2 and HSP27 plays a key role in the synergistic effects. If our hypotheses are true, the reslutant knowledge will be advantage for cooperative diagnosis of EOC and may also provide potent novel targeted regulation of MDR, recurrence and metastasis. Understanding the mechanisms by which tumor cells are able to circumvent elimination by chemotherapeutic agents and adopt a more aggressive behavior will hopefully lead to the development of more efficacious chemotherapeutic protocols to eradicate ovarian cancer cells. AS APP2 may be the disease-specific expression of endothelial cell surface protein, we also attempt to verify APP2 is a new selective marker of lymphatic endothelium in ovarian cancer tissues, which has the potential for increasing the accuracy of detection and treatment of lymphatic invasion in primary tumors. The receptors of TMTP1 may be widely applied for this purpose to targeting prevents the recurrence and metastasis of EOC.

尽管手术以及更为有效联合化疗后，多数的上皮性卵巢癌患者最终仍复发并发生紫杉醇和铂类的耐药。寻找新的基因作为突破点用于逆转耐药性卵巢癌基因治疗、抑制其转移及复发，已成为研究热点。卵巢癌侵袭转移与耐药间关系的研究也逐渐受到关注。申请人在前期研究中已证实靶向多肽TMTP1对顺铂耐药和高转移潜能上皮性卵巢癌均具有很好的靶向性，并成功筛选到TMTP1的受体为APP2和HSP27。本课题以该受体为出发点，提出APP2可能是上皮性卵巢癌发病中一个新基因的科学假设，探讨APP2和HSP27是否在卵巢癌发病机制中起关键协同作用，为靶向调控卵巢癌的耐药与转移寻找到共同作用的靶点。同时试图证实APP2为卵巢癌组织新生淋巴管内皮细胞的一种特异性标记物以及建立多种淋巴管模型用于探讨靶向多肽受体是否与肿瘤的淋巴管生成有关，可为卵巢癌的淋巴道转移的诊治提供新的思路。从而通过对靶向受体的调控实现上皮性卵巢癌的靶向阻遏。

项目背景：寻找新的基因作为突破点用于逆转耐药性卵巢癌基因治疗、抑制其转移及复发，已成为研究热点。申请人在前期研究中已证实靶向多肽TMTP1对顺铂耐药和高转移潜能上皮性卵巢癌均具有很好的靶向性，并成功筛选到TMTP1的受体为APP2和HSP27。主要研究内容：提出APP2可能是上皮性卵巢癌发病中一个新基因的科学假设，探讨APP2和HSP27是否在卵巢癌发病机制中起关键协同作用，为靶向调控卵巢癌的耐药与转移寻找到共同作用的靶点。重要结果、关键数据及其科学意义：我们已证实TMTP1的受体是APP2和HSP27以及PI3K/HSP27信号通路负性调节TMTP1所诱导肿瘤细胞的凋亡。APP2在不同转移潜能的肿瘤组织有表达差异性，提示APP2可能肿瘤转移明侵袭相关的一个新标志物；同时APP2能很好的识别卵巢癌耐药细胞，这为靶向阻遏卵巢癌的耐药提供了切实可行的方法，并在斑马鱼试验中得到证实；另一方面，我们筛选到一个与卵巢癌侵袭与转移相关的新靶点。TRA2B是一种RNA结合蛋白，通过RNA全基因测序，观察到几种不同的RNA剪切事件。TRA2B可以引起多种基因的表达上调或下调（CYP61，FN1，HMGA2，CTGF，ASAP3，ERBB3，MAP2K6，NPNT，IL-6，IL-11，JUN和ODC1）。由RNA测序数据量化的基因表达结果，证实了DEGs在肿瘤生长过程和转移中的表达水平有明显异常的变化。通过三个不同干扰片段（siRNA405，siRNA581以及siRNA798）对蛋白表达不同的抑制实验，发现干扰48h时，作用非常明显。以siRNA798最为明显。TRA2BshRNA和TRA2BsiRNA对不同的卵巢癌细胞（A2780和HO8910）的TRA2B蛋白有明显的抑制表达。与对照组相比，TRA2BshRNA和TRA2BsiRNA798对A2780和HO8910细胞内的CYP61，FN1，HMGA2和CTGF表达有明显抑制作用。同时可以抑制肿瘤肿瘤细胞的增殖和迁移能力，并促进肿瘤细胞的凋亡。可为卵巢癌的淋巴道转移的诊治提供新的思路。从而通过对靶向受体的调控实现上皮性卵巢癌的靶向阻遏。