肾癌新基因GSTM3调控区SNP与肾癌易感性及预后的关系

Renal cell carcinoma (RCC) is the major malignant tumor in urological system. The incidence of RCC increases constantly these years.The cause of RCC is unclear yet, there is no biomarkers for early prediction and diagnosis.Our previous studies have detected that Glutathione s transferase 3 (GSTM3) is a novel tumor suppressor gene in RCC. We aim to investigate the association among the GSTM3 functional polymorphisms in regulation regions, the interaction with environmental factors and the susceptibility to RCC using 500 cases and 1000 healthy controls in Chinese Han population. The following steps will be conducted: first, select the functional single nucleotide polymorphism (SNP) sites in GSTM3 gene regulation regions, detect the distributions of these SNP sites and analysis genotype frequencies, alleles and haplotypes in genome DNA of the control group; second, construct reporter plasmid containing different polymorphims, transfect them into the human embryo kidney 293 cells, explore their functional activities using dual-luciferase activity detection, real time quantitative PCR and etc, analyze interactions between the environmental factors and the valid functional polymorphism sites; lastly, determinate the relationship among the valid polymorphisms of GSTM3 gene, environmental factors and the occurance and prognosis according to the environmental relevant information. Our study aims to providing new theory and method for early prevention of RCC.

肾细胞癌（RCC）是泌尿系统主要恶性肿瘤，近年来其发病率迅速上升。目前肾癌发病原因不明，没有早期预测和诊断标志。我们前期研究发现谷胱甘肽转移酶（Glutathione s transferase 3, GSTM3）是肾癌抑制新基因。本课题拟应用500例病例和1000例正常对照，研究GSTM3功能snp位点、环境因素及其交互作用与肾癌易患性的关系：首先筛选GSTM3基因调控区（启动子区、miRNA靶序列区）可能有生物学意义的单核苷酸多态（SNP）位点，检测其在对照人群中的分布；其次通过单倍型分析、双荧光素酶报告基因系统检测等方法探讨碱基变异对GSTM3表达和功能的影响，确定出有意义的SNP，并研究有意义位点与环境因素的交互作用；最后结合正常人群的环境因素和基础疾病等相关信息，探讨GSTM3的功能多态性与环境因素、与肾癌发生和预后的关系，为肾癌早期预防提供新理论、新方法。

肾细胞癌（RCC）是泌尿系统主要恶性肿瘤，近年来其发病率迅速上升。目前肾癌发病原因不明，没有早期预测和诊断标志。我们前期研究发现谷胱甘肽转移酶（Glutathione s transferase 3, GSTM3）是肾癌抑制新基因。本课题应用329例ccRCC病例和420例正常对照，研究GSTM3基因调控区如启动子区和3‘UTR的SNP位点、环境因素及其交互作用与肾癌易患性的关系：结果发现启动子区SNPrs1332018和3‘UTR的SNP rs1055259与肾癌易感性显著相关，其中rs1332018显著影响RCC患者的预后。rs1055259的变异G等位基因干扰miR-556与GSTM3的3'UTR结合，从而增强GSTM3表达，并降低ROS活性，增高还原性巯基水平，最终抑制了ccRCC细胞的增殖和侵袭。 本课题通过探讨ccRCC患者群体中基因调控区功能SNP的发生及分布、基因型频率；然后进行单倍型分析、双荧光素酶报告基因系统检测、免疫荧光等分析方法研究SNP对基因表达和ccRCC发生和发展的影响，确定出有生物学意义的SNP及潜在功能；最后结合正常人群的环境因素和基础疾病等相关信息，探讨GSTM3基因的功能多态性与肾细胞癌发生和预后的关系，为肾癌早期预防提供新理论、新方法。