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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 摘要： 乳腺癌是40岁至55岁女性癌症相关死亡的主要原因，也是美国女性癌症相关死亡的第二大原因。暴露在环境、职业环境和饮食中的致癌物质被认为是造成差异的原因。流行病学研究表明，乳腺癌和吸烟有正相关关系。苯并(A)芘(B(A)P)是香烟烟雾的主要成分，已被证明会导致肺癌、肝癌、胰腺癌、结肠癌和乳腺癌。B(A)P被生物转化为反应性代谢产物，从而产生DNA加合物和DNA链断裂。此外，B(A)P已被证明可以将人乳腺上皮细胞(MCF-10A)从正常表型转变为癌变表型。一些OSCs二烯丙基硫化物(DAS)、二烯丙基二硫(DADS)和二烯丙基三硫(DATS)已被证明通过改变生物转化来预防癌症。我们推测，这些OSCs通过改变B(A)P的生物转化，从而阻止B(A)P DNA加合物的产生和细胞转化相关基因的表达，从而阻止正常人类乳腺上皮细胞向癌变表型的转化。为了验证这一假设，我们将首先用B(A)P和/或OSCs处理MCF-10A细胞，并确定是否形成反应性代谢物。其次，我们将通过彗星分析来确定苯并(A)P是否导致DNA链断裂，并通过P32后标记来确定DNA加合物，以及OSCs是否可以防止这种DNA损伤并随着时间的推移促进DNA修复。接下来，我们将确定B(A)P将正常细胞转化为癌变表型的能力，以及OSCs通过反复用B(A)P和OSCs处理细胞来抑制这种转化的能力。软琼脂上菌落的形成将表明转化细胞的存在。将这些转化的细胞移植到裸鼠体内，肿瘤的发展将证实肿瘤转化。最后，我们将证明B(A)P和/或OSCs将改变与转化和增殖相关的基因的表达。基因的表达将通过实时定量聚合酶链式反应阵列和Western Blot分析进行确认。这项研究的结果将有助于阐明B(A)P导致乳腺癌的机制，以及口腔干细胞如何预防这种癌症。最终，将为这项研究开发更好的化学预防方案和化疗化合物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.