肿瘤相关巨噬细胞外分泌QSOX1分子在原发性肝细胞癌发生发展中的作用及其机制研究

Quiescin sulfhydryl oxidase 1 (QSOX1) belongs to a member of FAD-linked sulfhydryl oxidase family, which plays an important role in modulating cell proliferation and apoptosis. Our previous study indicated that QSOX1 was the differential key molecule secreted from alternatively activated tumor associated microphage (M2) in tumor environment, which continue to rise during the HCC pathological stages. However, the specific mechanism deserves further investigation. In this research, by the way of constructing the overexpressed QSOX1 macrophage cell lines and quantitative proteomic technique, the QSOX1 related differential protein profiling would be established. The key molecules screened out will be analyzed for their folding status, spatial structures and their functions such as matrix remodeling and cell growth by the analytic method of protein disulfide bond. Using the nude mice HCC model and the cell singaling chips analysis, the possible sub network between QSOX1 related cell signaling and post translational modification of signal proteins will be established. Finally, we try to verify these relationships through detecting the expression of QSOX1 in HCC patients' serum. The purpose of this study was to illustrate the significance of QSOX1 in carcinogenesis and explore the novel biomarker for the early diagnosis and treatment of HCC.

巯基氧化酶1(QSOX1)是一种黄素依赖型巯基氧化酶家族蛋白,直接参与调控细胞增殖、凋亡等生物学进程。前期研究发现QSOX1在替代激活的肿瘤相关巨噬细胞(M2)分泌的上清液中高表达，且在正常人、肝炎、肝硬化、肝癌病人血清中的表达呈逐步递增趋势，但其作用机制尚末阐明。据此提出假说：QSOX1促进肝癌的发生发展。本课题拟通过构建QSOX1高表达巨噬细胞与肝癌细胞的共培养体系，采用定量蛋白质组学技术，建立QSOX1相关差异蛋白表达谱，运用蛋白质二硫键解析方法，探索重要差异蛋白的空间结构及其对细胞间基质重构、蛋白功能、细胞生长的影响；拟构建裸鼠肝癌模型，运用信号通路芯片，分析QSOX1可能的信号通路网络及信号分子翻译后修饰状态；再用临床血清标本，验证QSOX1与肝癌的潜在关系。本研究旨在揭示QSOX1在肝癌发生发展中的作用，以期发现肝癌新的分子标志物，为肝癌的早诊早治提供新思路。

巯基氧化酶 1(QSOX1)是一种黄素依赖型巯基氧化酶家族蛋白,直接参与调控细胞增殖、凋亡等生物学进程。前期研究发现 QSOX1 在替代激活的肿瘤相关巨噬细胞(M2)分泌的上清液中高表达，且在正常人、肝炎、肝硬化、肝癌病人血清中的表达呈逐步递增趋，但其作用机制尚末阐明。据此提出假说：QSOX1促进肝癌的发生发展。本课题首先通过干扰QSOX1基因在SMMC-7721细胞中的表达，检测其对肝癌细胞生物学功能的影响，发现干扰QSOX1基因在SMMC-7721细胞中的表达，能降低SMMC-7721肝癌细胞的增殖能力；克隆形成实验亦表明肿瘤细胞成瘤能力明显减弱。通过构建 QSOX1高表达巨噬细胞，发现高表达QSOX1巨噬细胞的条件培养基可增加SMMC-7721肝癌细胞的增殖、迁移和侵袭能力；通过构建过表达QSOX1巨噬细胞与肝癌细胞的共培养体系，采用定量蛋白质组学技术，建立 QSOX1 相关差异蛋白表达谱；进一步构建裸鼠皮下成瘤模型，探索过表达QSOX1巨噬细胞在肝癌发生发展中的作用；最后运用临床血清标本，验证 QSOX1 与肝癌的潜在关系。本研究揭示了 QSOX1 在肝癌发生发展中的作用，并为肝癌的诊断提供了新的分子标志物。