THE PREVENTION OF B(A)P INDUCED CELL TRANSFORMATION BY ORGANOSULFUR COMPOUNDS

有机硫化合物预防 B(A)P 诱导的细胞转化

• 批准号: 8357115
• 负责人: Selina Faith Darling-Reed
• 金额: $ 9.36万
• 依托单位: FLORIDA AGRICULTURAL AND MECHANICAL UNIV
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357115
• 关键词: Agar; Age; Benzo(a)pyrene; Breast; Cancer Etiology; Cancerous; Carcinogens; Cells; Cessation of life; Chemopreventive Agent; Colon Carcinoma; Comet Assay; DNA Adducts; DNA Damage; DNA Repair; DNA strand break; Development; Diet; Environment; Epidemiologic Studies; Epithelial Cells; Exposure to; Funding; Gene Expression; Grant; Human; Implant; Label; Lung; Malignant Neoplasms; Malignant neoplasm of liver; Malignant neoplasm of pancreas; Metabolic Biotransformation; National Center for Research Resources; Neoplastic Cell Transformation; Normal Cell; Nude Mice; Occupational; Pharmacologic Substance; Phenotype; Phosphorus 32; Prevention; Principal Investigator; Production; Regimen; Research; Research Infrastructure; Resources; Source; Testing; Time; United States; United States National Institutes of Health; Western Blotting; Woman; abstracting; allyl sulfide; benzo(a)pyrene-DNA adduct; cell transformation; cigarette smoking; cost; diallyl disulfide; diallyl trisulfide; malignant breast neoplasm; prevent; tumor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Abstract: Breast cancer is the leading cause of cancer related death in women between the ages of 40 and 55 and the second leading cause of cancer related death in women in the United States. Exposure to carcinogens in the environment, occupational setting, and diet are believed to be the reason for the discrepancy. Epidemiological studies demonstrate a positive association with breast cancer and cigarette smoke. Benzo(a)pyrene (B(a)P), a major component of cigarette smoke, has been shown to cause lung, liver, pancreatic, colon and breast cancer. B(a)P is biotransformed into reactive metabolites that produces DNA adducts and DNA strand breaks. Furthermore, B(a)P has been shown to transform human breast epithelial cells (MCF-10A)from a normal to a cancerous phenotype. Several OSCs Diallyl Sulfide (DAS), Diallyl Disulfide (DADS) and Diallyl Trisulfide (DATS) have been shown to prevent cancer by altering biotransformation. We hypothesize that these OSCs will prevent the transformation of normal human breast epithelial cells to a cancerous phenotype by altering the biotransformation of B(a)P thus preventing the production of B(a)P DNA adducts and the expression of genes involved in cell transformation. To test this hypothesis first we will treat MCF-10A cells with B(a)P and/or OSCs and determine if reactive metabolites are formed. Secondly, we will determine if B(a)P cause DNA strand breaks by comet assay analysis and DNA adducts via P32 Post labeling and if OSCs can prevent this DNA damage and enhance DNA repair over time. Next we will determine the ability of B(a)P to transform normal cell into a cancerous phenotype and the ability of OSCs to inhibit this transformation by treating the cells repeatedly with B(a)P and OSCs. The formation of colonies on soft agar will indicate the presence of transformed cells. Tumor development in nude mice implanted with these transformed cells will confirm neoplastic transformation. Finally, we will demonstrate that B(a)P and/or OSCs will alter the expression of genes associated with transformation and proliferation. Gene expression will be determined by Real Time PCR Array and confirmed by Western Blot Analysis. Results from this study will help elucidate the mechanism by which B(a)P causes breast cancer and how OSCs may prevent this cancer. Ultimately, better chemopreventive regimens and chemotherapeutic compounds would be developed for this research.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其他NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， NCRR赠款不直接向子项目或子项目工作人员提供资金。 摘要： 在美国，乳腺癌是40至55岁女性癌症相关死亡的主要原因，也是女性癌症相关死亡的第二大原因。 暴露于环境中的致癌物质，职业环境和饮食被认为是差异的原因。流行病学研究表明，吸烟与乳腺癌呈正相关。 苯并（a）芘（B（a）P）是香烟烟雾的主要成分，已被证明可导致肺癌、肝癌、胰腺癌、结肠癌和乳腺癌。B（a）P被生物转化为产生DNA加合物和DNA链断裂的反应性代谢物。 此外，已显示B（a）P将人乳腺上皮细胞（MCF-10 A）从正常表型转化为癌性表型。 几种OSC二烯丙基硫化物（DAS），二烯丙基二硫化物（DADS）和二烯丙基三硫化物（DATS）已被证明可以通过改变生物转化来预防癌症。我们假设这些OSC将通过改变B（a）P的生物转化从而阻止B（a）P DNA加合物的产生和参与细胞转化的基因的表达来阻止正常人乳腺上皮细胞转化为癌性表型。为了检验这一假设，我们首先用B（a）P和/或OSC处理MCF-10 A细胞，并确定是否形成反应性代谢物。 其次，我们将通过彗星试验分析确定B（a）P是否引起DNA链断裂和通过P32后标记的DNA加合物，以及OSC是否可以防止这种DNA损伤并随着时间的推移增强DNA修复。 接下来，我们将确定B（a）P将正常细胞转化为癌性表型的能力，以及通过用B（a）P和OSC重复处理细胞来抑制这种转化的能力。 软琼脂上菌落的形成表明存在转化细胞。 移植这些转化细胞的裸鼠中的肿瘤发展将证实肿瘤转化。 最后，我们将证明，B（a）P和/或OSC将改变与转化和增殖相关的基因的表达。 基因表达将通过真实的时间PCR阵列测定，并通过蛋白质印迹分析确认。 这项研究的结果将有助于阐明B（a）P导致乳腺癌的机制以及OSCs如何预防这种癌症。最终，更好的化学预防方案和化疗化合物将开发这项研究。