Lipid-Peroxidation Induced Cyclic Adducts in Hepatocarcinogenesis

脂质过氧化诱导的环状加合物在肝癌发生中的作用

• 批准号: 7656102
• 负责人: FUNG-LUNG CHUNG
• 金额: $ 46.38万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7656102
• 关键词: Acrolein; Affect; Aldehydes; Animal Model; Animals; Antioxidants; Base Excision Repairs; Binding; Binding Sites; Biological; Biological Assay; Carcinogens; Cells; Chemopreventive Agent; Chronic; Cinnamon - dietary; Classification; Codon Nucleotides; Copper; DNA; DNA Damage; DNA lesion; Data; Defect; Deoxyguanosine; Development; Diagnostic Neoplasm Staging; Fatty Liver; Frequencies; Goals; Green tea; Guanine; Harvest; Hepatocarcinogenesis; Human; In Vitro; Inbred LEC Rats; Incidence; Individual; Knockout Mice; Lesion; Life; Ligation; Light; Link; Lipid Peroxidation; Liver; Liver neoplasms; Long-Evans Rats; Lung; Malignant Neoplasms; Malignant neoplasm of liver; Maps; Mediating; Modeling; Monitor; Mus; Mutate; Mutation; Mutation Spectra; Nucleotide Excision Repair; Obesity; Organ; Oxidative Stress; Pathway interactions; Pattern; Polyphenon E; Polyunsaturated Fatty Acids; Primary carcinoma of the liver cells; Risk Factors; Rodent; Role; Skin Carcinogenesis; Slice; Staging; TP53 gene; Thioctic Acid; Thymine; Tissues; Transgenic Organisms; Tumor Tissue; Tumor stage; UV Radiation Exposure; UV induced; Vitamin E; XPA gene; adduct; base; carcinogenesis; human DNA; human tissue; in vivo; oxidation; repaired; tool; tumor; tumorigenesis

DESCRIPTION (provided by applicant): Cyclic 1,N2-propanodeoxyguanosine adducts of acrolein (Acr-dG) and trans-4-hydroxy-2-nonenal (HNE-dG) derived from lipid peroxidation have been detected as endogenous DNA lesions in rodent and human tissues. However, their roles in carcinogenesis are still not clearly understood. Recent in vitro studies in cultured human cells and in DNA fragments containing the human p53 gene have shown that Acr and HNE can bind to the p53 gene preferentially at guanines in a sequence-selective manner and that the binding patterns are similar to p53 mutation hotspots observed in certain human cancers, suggesting the possible roles of Acr-dG and HNE-dG in carcinogenesis. However, no in vivo studies have been conducted to directly examine cyclic adducts in tissues where tumors develop in a relevant animal tumorigenesis model. These studies are important because they provide strong evidence directly linking adducts with mutations and tumor formation in the target tissue. In this application, we propose to use genetically defected Long Evans Cinnamon (LEC) rats and transgenic XPA-deficient (XPA-/-) mice, both of which are highly prone to spontaneous liver cancer without carcinogen treatment, to investigate the roles of Acr-dG and HNE-dG as endogenous DNA lesions in carcinogenesis. LEC rats are inflicted with heightened lipid peroxidation in the liver as a result of abnormal copper accumulation, whereas XPA(-/-) mice are deficient in nucleotide excision repair (NER) which is important for removing Acr- and HNE-dG adducts. Our hypothesis is that the formation of the cyclic adducts will be increased and these adducts will accumulate in the liver DNA of these animals compared with the wild-type Long Evans (LE) rats and XPA(+/+) mice and that the increased formation of these adducts underlies the mechanisms for liver carcinogenesis. We will carry out the following aims to examine this hypothesis. In Aim 1, we will conduct tumor bioassays in LEC rats and XPA(-/-) mice and their normal strains and study throughout their lives the relationships of cyclic adducts in the liver DNA with liver carcinogenesis at different stages; In Aim 2, we will map Acr and HNE adduct binding sites in the p53 gene of the liver DNA from LEC rats and XPA (-/-) mice during the tumor bioassays and compare their binding sites with the p53 mutational spectra of the liver tumors harvested at the termination of bioassays. In Aim 3, we will investigate the effects of antioxidants on the formation of cyclic adducts in the liver DNA and on the development of spontaneous liver cancers in these animals. The data generated from these studies will help us to understand the roles of endogenous cyclic Acr-dG and HNE-dG adducts from lipid peroxidation in hepatocarcinogenesis.

描述(申请人提供)：在啮齿动物和人类组织中检测到源于脂质过氧化的环状1，2-丙基脱氧鸟苷加合物(ACR-DG)和反式-4-羟基-2-壬烯醛(HNE-DG)是内源性DNA损伤。然而，它们在肿瘤发生中的作用仍不清楚。最近在体外培养的人细胞和含有人P53基因的DNA片段中的研究表明，ACR和HNE能以序列选择性的方式优先与P53基因结合，结合模式与某些人类癌症中观察到的P53突变热点相似，提示ACR-DG和HNE-DG在肿瘤发生中可能发挥作用。然而，还没有进行体内研究，以直接检查在相关动物肿瘤发生模型中发生肿瘤的组织中的环状加合物。这些研究很重要，因为它们提供了强有力的证据，直接将加合物与靶组织中的突变和肿瘤形成联系起来。在这一应用中，我们建议使用基因缺陷的Long Evans Cinnamon(LEC)大鼠和转基因XPA缺陷(XPA-/-)小鼠，这两种小鼠在没有致癌剂治疗的情况下都很容易发生自发性肝癌，以探讨ACR-DG和HNE-DG作为内源性DNA损伤在肿瘤发生中的作用。由于铜的异常蓄积，LEC大鼠肝脏脂质过氧化加剧，而XPA(-/-)小鼠缺乏核苷酸切除修复(NER)，NER是清除ACR-和HNE-DG加合物的重要因素。我们的假设是，与野生型Long Evans(LE)大鼠和XPA(+/+)小鼠相比，这些动物的环状加合物的形成将增加，这些加合物将在肝脏DNA中积聚，这些加合物的形成增加是肝癌发生机制的基础。我们将执行以下目的来检验这一假说。在目标1中，我们将在LEC大鼠和XPA(-/-)小鼠及其正常品系中进行肿瘤生物检测，并在其一生中研究肝DNA中的环状加合物与不同阶段肝癌发生的关系；在目标2中，我们将在肿瘤生物检测期间定位LEC大鼠和XPA(-/-)小鼠肝DNA P53基因中的ACR和HNE加合物结合位点，并将它们的结合位点与生物检测结束时收获的肝肿瘤的P53突变谱进行比较。在目标3中，我们将研究抗氧化剂对肝DNA中环状加合物形成的影响，以及对这些动物自发性肝癌发生的影响。这些研究产生的数据将有助于我们理解内源性环状ACR-DG和HNE-DG脂质过氧化加合物在肝癌发生中的作用。