"蛋白组筛查- - 功能分析-药物靶标"技术链分析RACK1蛋白群组在口腔黏膜癌变诊断及治疗中的潜能

项目名称： "蛋白组筛查- - 功能分析-药物靶标"技术链分析RACK1蛋白群组在口腔黏膜癌变诊断及治疗中的潜能

项目编号： No.30801294

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 王智

作者单位： 四川大学

项目金额： 21万元

中文摘要： 口腔癌变关键分子事件是揭秘癌变机制，开发靶向药物的核心。本课题拟在前期蛋白组学在粘膜癌变中新发现的候选标记物蛋白激酶K受体1（RACK1）为核心，集中开展以下四个方面的研究工作：①#25506;求RACK1蛋白表达在临床诊断预后判断中的价值；②#26500;建壳聚糖－PCEC纳米颗粒包被shRNA重组质粒复合转染系统，测定转染效率和细胞毒性。③#21033;用此复合转染系统沉默RACK1基因，体外试验观察RACK1基因沉默后口腔鳞癌细胞增殖、凋亡、黏附、侵袭、迁移变化；⑤#21033;用此复合转染系统沉默RACK1基因，建立荷瘤鼠模型，体内试验明确RACK1基因沉默后抗肿瘤生长活性。研究显示RACK1随异常增生程度增加而表达递增，可作为临床诊断分子标记物； RACK1蛋白转移组高表达，及与Ki67比较结果均显示其是良好的肿瘤预后判定因子；RACK1基因表达量下降可致口腔鳞癌细胞凋亡增加，增殖，侵袭黏附降低，RACK1可调节口腔鳞癌凋亡途径，可作为候选药物靶标。本项目以功能蛋白组学研究为基础，整合RNA干扰及纳米技术，从临床样本到细胞分子水平，从体外到体内确立RACK1及相关蛋白功能，形成"蛋白组筛查- - 功能分析-药物靶标"技术链。

中文关键词： 口腔鳞癌；壳聚糖-PCEC纳米颗粒包被shRNA质粒转染系统；预后；RNA干扰；靶向治疗

英文摘要： The research project targeting to RACK1 will focus on four aspects:first, to establish the RACK1 related protein chain and study the signal transduction of carcinogenesis by microarray;second,to investigate the role of RACK1 in diagnosis and prognosis evaluation;third,to establish the chitoson-PCEC nanopartical packing shRNA transfection system and evaluate trandfection efficiency and cytotoxity;fourth,to use this transfection system to silence RACK1 gene and observe the apoptosis,invasion and migration phenotype in vitro.fifth, by using the transfection system and the xenografts model,to observe the tumor growth. Immunohistochemical analysis of tissues exhibited an increased expression of RACK1 with increasing of pathological classification. Following analysis with epidemic factors showed that up-regulation of RACK1 was related to relatively poor prognosis of OSCC. Silencing RNA interference specifically on RACK1 could initiate apoptosis and inhibit proliferation,migration,invasion of OSCC cells.In all ,the function role of RACK1 was established in our project by combining proteomic technique with RNA interference and nanoparticle from clinical to molecular level,from in vitro to in vivo.With this, a technique chain"proteomic screening- - functional analysis-target therapy" was preliminarily set up.

英文关键词： OSCC;chitoson-PCEC nanopartical packing shRNA transfection system; RNA inteference;target therapy