代谢型谷氨酸受体4在人骨肉瘤发生发展中的作用机制研究

项目名称： 代谢型谷氨酸受体4在人骨肉瘤发生发展中的作用机制研究

项目编号： No.81460407

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 贺茂林

作者单位： 广西医科大学

项目金额： 47万元

中文摘要： 骨肉瘤是最常见的原发恶性骨肿瘤。国外的全基因组关联分析发现，mGluR4基因的rs1906953与骨肉瘤相关，我们的前期研究也发现，mGluR4在骨肉瘤中的表达高于正常骨组织，且具有mGluR4高表达骨肉瘤患者的预后较差。我们拟采用甲基化特异性的PCR法，对mGluR4基因进行启动子甲基化检测；采用PCR等位基因分型检测mGluR4基因在骨肉瘤患者和正常人群中的多态性；筛选出具有mGluR4高表达的骨肉瘤细胞系后，分别采用mGluR4的激动剂、抑制剂以及构建的RNAi慢病毒表达载体，对骨肉瘤细胞进行处理，观察细胞周期、增殖活性、侵袭能力和裸鼠成瘤能力等生物学行为的变化情况；最后，我们还将采用实时荧光定量PCR阵列分析技术探索mGluR4基因对骨肉瘤产生影响的信号通路和作用靶点，并利用免疫蛋白印迹、免疫荧光染色及免疫共沉淀等技术进行验证，从分子生物学上探讨人骨肉瘤发生、发展的作用机制。

中文关键词： 骨肉瘤；RNA干扰；PCR阵列；代谢型谷氨酸受体4；单核苷酸多态性

英文摘要： Osteosarcoma is the most common primary malignant bone tumor. The foreigner made genome-wide association analysis study and found that the rs1906953 of mGluR4 gene is related to osteosarcoma; also, our previous research has found that the expression of mGluR4 in osteosarcoma is significantly higher than that of the normal bone tissue, and patients with osteosarcoma of high mGluR4 expression has poor prognosis. On this basis, to explore the functions of mGluR4 in the occurence and development of osteosarcoma, the methylation-specific PCR method will be used to make methylation inspection to the promoters;the method of PCR allele classification will be used to test for the polymorphism of mGluR4 gene in both patients with osteosarcoma and in normal people;after screening out the osteosarcoma cell line with high expression of mGluR4, we respectively use the agonist and inhibitor of mGluR4, and use the established RNAi lentiviral expression vector to treat the osteosarcoma cells, investigate the changes on the cell cycle, proliferative activity, invasion ability, tumor formation capacity in nude mice and other biological behaviors; finally, we will use real-time fluorescent quantitative PCR array analysis technology to explore the signaling pathways and targets of the influence of mGluR4 gene on osteosarcoma,we propose to use Western Blot、Immunofluorescence and Co-Immunoprecipitation techniques to identify it,and to explore the mechanism of the occurrence and development of osteosarcoma from molecular biology.

英文关键词： Osteosarcoma;RNA interference;PCR Array;Metabotropic Glutamate Receptor 4;Single Nucleotide Polymorphism