Quinone Oxidoreductases and Mammary Toxicity/Carcinogenenicity

醌氧化还原酶和乳腺毒性/致癌性

• 批准号: 8662266
• 负责人: Anil Kumar Jaiswal
• 金额: $ 34.19万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8662266
• 关键词: Affect; Anthracenes; Apoptosis; BRCA1 gene; BRCA2 gene; Biological Assay; Biological Markers; Breast Cancer Prevention; Catechol Estrogens; Catechols; Cause of Death; Chemicals; DNA Repair; Development; Differentiation and Growth; Drug Metabolic Detoxication; Duct (organ) structure; Endogenous Factors; Estradiol; Estrogens; Frequencies; Gelatinase B; Gene Family; Genes; Genetic; Goals; Histology; Human; Immunoprecipitation; In Vitro; Individual; Intraductal Hyperplasia; Laboratories; Liver; Lobule; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Mammary Gland Parenchyma; Mammary Neoplasms; Mammary Tumorigenesis; Mammary gland; Mass Spectrum Analysis; Mediating; Metabolic; Metabolic Activation; Metabolism; Metastatic Neoplasm to the Breast; Milk; Molecular; Mouse Mammary Tumor Virus; Mus; Mutation; NAD(P)H dehydrogenase (quinone) 1, human; NQO1 gene; Neoplasm Metastasis; Organ; Phosphorylation; Play; Predisposition; Proteasome Binding; Proteins; Quinone Metabolism; Quinones; Radiation; Risk; Risk Factors; Role; Skin Cancer; Spleen; Stress; Testing; Time; Tissues; Toxic effect; Transfection; Wild Type Mouse; Woman; carcinogenicity; dimethylbenzanthracene; in vivo; malignant breast neoplasm; member; multicatalytic endopeptidase complex; non-genetic; protein degradation; public health relevance; research study; tumor

DESCRIPTION (provided by applicant): Mammary cancer is the leading cause of death among women. About 5-10% is the familial cases, which are attributed to deleterious mutations in BRCA1 and BRCA2. The majority of the remaining breast cancer cases are also suspected to contain genetic component that affects BRCA1/BRCA2. Preliminary studies revealed that quinone oxidoreductases (NQO1 and NQO2) might protect against mammary toxicity, carcinogenicity and metastasis. Disruption of NQO1 gene in mice led to mammary gland intraductal hyperplasia and DMBA-induced mammary tumors that metastasized to lung, liver and spleen. NQO2 is highly homologous to NQO1 and is also expected to protect against mammary cancer. Preliminary studies also showed that chemical/radiation stress responsive NQO1 and NQO2 regulate stability of BRCA1 against 20S proteasomal degradation that contributes to protection against DMBA-induced mammary cancer. In addition, NQO1 and NQO2 control stability of C/EBPa that suppresses matrix metalloproteinase9 (MMP9) and protect against metastasis. The overall goal of the current studies is to investigate the hypothesis that NQO1 and NQO2 are endogenous factors in protection against mammary toxicity, carcinogenicity and metastasis. Three aims are planned. Aim 1 will test the hypothesis that NQO1 and NQO2 are biomarkers of susceptibility to DMBA-induced and spontaneous mammary cancer and metastasis. In addition, this aim will also investigate the hypothesis that NQO1 and NQO2 metabolic detoxification of catechol quinones and/or control of factors in DNA repair, growth/differentiation and apoptosis protect against mammary cancer. Wild type, NQO1-/-, NQO1+/- and NQO2-/- mice exposed to DMBA will be analyzed for mammary tumor frequency/multiplicity and metastasis. NQO1-/- and NQO2-/- mice will also be crossed with MMTV-Neu-p53 mice and assessed for increase in frequency and multiplicity and decrease in time of spontaneous mammary tumor development and metastasis. In addition, mice exposed to DMBA or estradiol will be analyzed for metabolic activation/detoxification of catechol estrogen quinones and key molecular changes in NQO1-/- and NQO2-/- mice leading to increased susceptibility to DMBA-induced mammary cancer/metastasis, as compared with wild type mice. Aim 2 will determine mechanism of NQO1 and NQO2 control of stability of BRCA1 against 20S proteasomal degradation leading to protection against mammary cancer. Western, immunoprecipitation, transfection, protein degradation, domains interaction, and phosphorylation assays will study NQO1/NQO2 control of BRCA1/BRCA2 stability against 20S degradation. Stress induced gain/loss, of NQO1/NQO2 and 20S proteasome interactions with BRCA1/BRCA2 will be followed. Aim 3 will elucidate the mechanism of NQO1 and NQO2 control of MMP9 protein leading to protection against mammary cancer metastasis. We will test the hypothesis that NQO1 and NQO2 stabilization of C/EBPa against 20S proteasomal degradation leads to suppression of MMP9 with implications in protection against metastasis. Studies are significant since human individuals carry deleterious mutations in NQO1 and NQO2 and are deficient in these proteins.

描述（由申请人提供）：乳腺癌是女性死亡的主要原因。大约5-10%是家族性病例，归因于BRCA1和BRCA2的有害突变。其余大多数乳腺癌病例也被怀疑含有影响 BRCA1/BRCA2 的遗传成分。初步研究表明，醌氧化还原酶（NQO1 和 NQO2）可能可以防止乳腺毒性、致癌性和转移。小鼠中 NQO1 基因的破坏导致乳腺导管内增生和 DMBA 诱导的乳腺肿瘤转移到肺、肝和脾。 NQO2 与 NQO1 高度同源，也有望预防乳腺癌。初步研究还表明，化学/辐射应激反应性 NQO1 和 NQO2 调节 BRCA1 的稳定性，防止 20S 蛋白酶体降解，从而有助于预防 DMBA 诱导的乳腺癌。此外，NQO1和NQO2控制C/EBPα的稳定性，从而抑制基质金属蛋白酶9（MMP9）并防止转移。 当前研究的总体目标是调查 NQO1 和 NQO2 是预防乳腺毒性、致癌性和转移的内源性因素的假设。计划了三个目标。目标 1 将检验以下假设：NQO1 和 NQO2 是 DMBA 诱导的自发性乳腺癌和转移易感性的生物标志物。此外，该目标还将研究 NQO1 和 NQO2 儿茶酚醌代谢解毒和/或控制 DNA 修复、生长/分化和细胞凋亡因子可预防乳腺癌的假设。将分析暴露于 DMBA 的野生型、NQO1-/-、NQO1+/- 和 NQO2-/- 小鼠的乳腺肿瘤频率/多样性和转移。 NQO1-/-和NQO2-/-小鼠还将与MMTV-Neu-p53小鼠杂交，并评估自发乳腺肿瘤发生和转移的频率和多样性的增加以及时间的减少。此外，将分析暴露于 DMBA 或雌二醇的小鼠儿茶酚雌激素醌的代谢激活/解毒以及 NQO1-/- 和 NQO2-/- 小鼠中导致对 DMBA 诱导的乳腺癌/转移的易感性增加的关键分子变化，如与野生型小鼠相比。目标 2 将确定 NQO1 和 NQO2 控制 BRCA1 稳定性以防止 20S 蛋白酶体降解从而预防乳腺癌的机制。 Western、免疫沉淀、转染、蛋白质降解、结构域相互作用和磷酸化测定将研究 NQO1/NQO2 对 BRCA1/BRCA2 20S 降解稳定性的控制。将跟踪压力诱导的 NQO1/NQO2 和 20S 蛋白酶体与 BRCA1/BRCA2 相互作用的增益/损失。目标 3 将阐明 NQO1 和 NQO2 控制 MMP9 蛋白从而防止乳腺癌转移的机制。我们将测试以下假设：NQO1 和 NQO2 对 C/EBPα 的稳定性抵抗 20S 蛋白酶体降解，从而抑制 MMP9，从而对防止转移产生影响。研究意义重大，因为人类个体携带 NQO1 和 NQO2 有害突变，并且缺乏这些蛋白质。