胞外分泌型人巯基氧化酶1(Es-hQSOX1b)剪接体功能及其生物药学意义探索

项目名称： 胞外分泌型人巯基氧化酶1(Es-hQSOX1b)剪接体功能及其生物药学意义探索

项目编号： No.31300660

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 郑文云

作者单位： 华东理工大学

项目金额： 22万元

中文摘要： 人巯基氧化酶1(hQSOX1)是多细胞生物中参与蛋白质二硫键形成的巯基氧化酶（QSOX）家族的一类酶。hQSOX1基因选择性剪接产生编码胞外分泌型（Es-hQSOX1b）和高尔基体膜结合型（Gb-hQSOX1a）两种mRNA分子。目前对胞内Gb-hQSOX1a的功能已确证，而对分泌于胞外基质的剪接体Es-hQSOX1b生理及病理功能尚不明了,最新研究推测Es-hQSOX1b可能是乳腺癌转好的一个新生物标志物。 本课题在前期制备高活性hQSOX1b和研究其不同突变体功能的基础上，拟利用免疫学、RNAi、荧光探针、细胞原位成像和细胞凋亡检测等技术，研究Es-hQSOX1b的抑制与过表达对正常细胞及癌细胞外基质主要相关蛋白结构的稳定性和酶对底物活性影响，及氧自由基产生而致细胞凋亡的作用，揭示Es-hQSOX1b对正常细胞和在癌发生及转移中的作用，发掘其生物药学价值，为癌症防治新药研发奠定基础。

中文关键词： QSOX；细胞外基质形成；蛋白质折叠；大肠杆菌细胞改造；CHO细胞改造

英文摘要： Sulphydryl oxidase QSOX1 is a kind of enzyme that can insert disulfide in reduced protein in multiceluler organisms. The QSOX1 gene can generate two mRNAs, secretory QSOX1b （Es-hQSOX1b）and Golgi-binding QSOX1a（Gb-hQSOX1a）. Some functions of Gb-hQSOX1a have been known, however, the functions of Es-hQSOX1b in phycological and pathycological states have been unclear. The latest research has implied that Es-hQSOX1b maybe is the biomarker of good prognosis of breast cancer. On the basis of precious work that the hQSOX1 with high activity have been prepared and the functions of mutants have been studied,the effect of Es-hQSOX1b overexpression and inhibition on stability and activity of extracellular natrix proteins, apoptosis induced by ROS will be studied by immunity, RNAi, fluorescent probe, in situ image and apoptosis detection. We hope to discover the function of Es-hQSOX1b in cancar generation and metastasis and form the basis for the discovery of new anticancer drug.

英文关键词： QSOX1；ECM formation；protein folding；E.coli modification；CHO modification