乳源六肽结合EEF1D逆转卵巢癌细胞耐药性及其分子机制

项目名称： 乳源六肽结合EEF1D逆转卵巢癌细胞耐药性及其分子机制

项目编号： No.81472448

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 秦宜德

作者单位： 安徽医科大学

项目金额： 72万元

中文摘要： 卵巢癌由于原发性与继发性耐药,是妇科治疗最为困难的恶性肿瘤。探索化疗耐药的分子机制, 提高化疗敏感性，对卵巢癌的治疗有重要意义。PGPIPN是一具有多种功能的乳源六肽。前期研究发现，PGPIPN显著增敏卵巢癌细胞株SKOV3的化疗，其结合靶蛋白是翻译延长因子1δ（EEF1D）。本项目利用耐药和敏感卵巢细胞株、化疗不敏感和敏感临床卵巢癌组织、SCID鼠荷瘤模型，研究PGPIPN通过结合和失活EEF1D逆转卵巢癌的药物抗性及其对耐药相关基因和信号途径影响，以阐明PGPIPN逆转卵巢癌细胞抗药及其机制。通过siRNA、2D电泳、LC-MS、IHC和cDNA芯片等，从分子水平研究EEF1D功能、耐药相关基因及信号途径，以明确耐药作用相关机制；用裸鼠种植瘤实验，从整体验证PGPIPN对EEF1D介导的耐药和相关机制。本项目通过探索乳源六肽逆转卵巢癌抗药机制，为卵巢癌治疗提供新思路、靶点、手段和药物。

中文关键词： C24_卵巢肿瘤；六肽；药物抗性；分子机制；靶蛋白

英文摘要： Ovarian cancer is one of the most serious malignant gynecological tumors. Due to primary and secondary development of multidrug resistance, the researches on decreasing chemoresistance, elucidating the molecular mechanism of resistance and improving chemotherapy sensitivity have important significance for the treatment of ovarian cancer. PGPIPN is a multifunctional hexapeptide derived from beta casein in bovine milk. Previous studies have found that PGPIPN significantly sensitized the chemotherapy of ovarian cancer cell line SKOV3, target protein of which is eukaryotic translation elongation factor 1 delta (EEF1D). This project will study that the effects of PGPIPN on the reversing chemoresistance of ovarian cancer cell, and what influence exerted by PGPIPN on genes associated multidrug resistance (MDR) and signaling pathways by binding and inactivating target protein-EEF1D. In this study, we will utilize resistant and sensitive ovarian cancer cell lines to drugs, clinical ovarian cancers insensitive to chemotherapy (progressive disease/stable disease, PD/SD) and sensitive to chemotherapy (complete response/partial response, CR/PR) in vitro, and the xenograft SCID mice model in vivo. We will study that PGPIPN contributes to reversing drug resistance of ovarian cancer mediated by EEF1D in the cellular level. By small interfering RNA (siRNA), two-dimensional electrophoresis, liquid chromatography-mass spectrometry (LC-MS), cDNA chip, real time PCR, immunohistochemistry (IHC) and Western Blot, bioinformatics (gene set enrichment, pathway, etc.) and other methods, we will probe EEF1D function as target of PGPIPN, genes related chemoresistance and signaling pathways，to clarify the reversal mechanism of ovarian cancer cell chemoresistance in the molecular level. These are validated through real-time quantitative PCR, western blot and luciferase reporter gene system. By the xenograft tumor SCID mice model, the about effects will be verified in the overall. By studying these mechanisms of PGPIPN reversing chemoresistance of ovarian cancer mediated by EEF1D, the project will provide new ideas, targets, methods and drugs for the therapy of ovarian cancer.

英文关键词： ovarian tumor;hexapeptide;chemoresistance;molecular mechanism;target protein