硫化氢在肝癌细胞乏氧辐射耐受中的作用机制研究

项目名称： 硫化氢在肝癌细胞乏氧辐射耐受中的作用机制研究

项目编号： No.31200631

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 生物物理、生化与生物分子学、生物力学与组织工程

项目作者： 张江虹

作者单位： 复旦大学

项目金额： 23万元

中文摘要： 乏氧细胞是造成肿瘤对放疗抵抗甚至愈后复发的重要原因之一，抑制SIRT1表达能提高肿瘤对照射和各种DNA损伤因素的敏感性。硫化氢在缺血缺氧性疾病中对细胞增殖和凋亡发挥着重要的调节作用。近期发现，H2S在乏氧肝癌细胞辐射损伤中起保护作用,本课题拟以不同p53状态的肝癌细胞为研究对象，通过shRNA转染建立SIRT1缺失的细胞系，结合siRNA干扰和化学抑制剂，采用体外集落形成实验、微核试验、流式细胞术、Western blot、Real time-PCR和DNA电泳等方法检测乏氧条件下内外源性硫化氢对肝癌细胞受照后细胞周期、凋亡、增殖和DNA损伤等；以p53、Caspase凋亡通路和ATP敏感钾通道为切入点，研究硫化氢在乏氧辐射耐受中的作用机制，研究H2S与SIRT1在乏氧辐射耐受诱导的p53凋亡通路中的相互关系。项目研究成果将为发现新的肿瘤治疗靶点、发展新的肿瘤治疗手段提供理论依据。

中文关键词： 乏氧；；硫化氢；；辐射抵抗;；辐射旁效应；；沉默信息调节因子1

英文摘要： It has been known that solid tumor always contain a hypoxic area due to chronic insufficiency in blood supply. The radioresistive hypoxic cells may result in unfavorable prognosis effects on tumor therapy.The sensibility of radiation and others DNA damage stresses can be promoted via the inhibition of SIRT1 (Sirtuin 1, SIRT1). Hydrogen sulfide(H2S), a new gasiform signaling molecule, which plays an important regulative action on cell proliferatioin and apoptosis in the ischemic and hypoxic diseases. Recently, We found that H2S played a protective effect for radiation damage on the hypoxic human hepatoma carcinoma cells. In this project, we will use hepatoma cell lines of HepG2 and Hep3B with different p53 phenotypes and their transfected cell lines without SIRT1 gene expression, named HepG2-SIRT1-shRNA and Hep3B-SIRT1-shRNA,treat the cells with siRNA and special inhibitors of target genes, measure irradiation damage with colony formation assay,micronuclei assay, flow cytometry assay, Western blot, Real time PCR, DNA electrophoresis, and investigate the radiation effect of endogenous and exogenous H2S on cell cycle, apoptosis, priliferation and DNA damage on hypoxic hepatoma cell. Moreover, the relation between H2S and SIRT1 and the role of H2S in hypoxia-induced radioresistivity including p53-apoptosis pathway,

英文关键词： Hypoxia;；Hydrogen sulfide;；Radiotolerance;；RIBE；Sirtuin 1