Carcinogen Metabolism Enzyme Expression in Buccal Cells

口腔细胞中致癌物代谢酶的表达

• 批准号: 6522774
• 负责人: SIMON D SPIVACK
• 金额: $ 19.06万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6522774
• 关键词: aromatic hydrocarbon receptor; bronchus; clinical research; cytochrome P450; enzyme induction /repression; estradiol; estrogen receptors; glutathione transferase; hormone related neoplasm /cancer; human subject; human tissue; liquid chromatography mass spectrometry; nicotine; nutrition aspect of cancer; oral mucosa; oxidoreductase; polymerase chain reaction; receptor expression; respiratory epithelium

DESCRIPTION (provided by applicant): This proposal's specific objective is to evaluate the validity of using gene expression in buccal cells to accurately reflect gene expression in human lung when both are tobacco smoke-exposed, as they are in smokers. The induction of genes encoding carcinogen metabolizing enzymes simultaneously in an easily-accessible surrogate tissue as well as the lung suggests the possibility of expression phenotyping smokers for bioactivating gene inducibility. If this carcinogen metabolism pathway is indeed relevant to lung cancer susceptibility, then a high phase I and low phase II enzyme inducibility phenotype might be hypothesized to confer risk for lung cancer. The power of studies on carcinogen-metabolizing gene induction in lung cancer to date have been limited by the number of subjects enrolled, as quality human lung tissues suitable for expression or activity assays are in short supply. Additionally, studies of coding sequence polymorphisms in these genes have presented very mixed results as to conferred risk for lung cancer. This study proposes to take advantage of newly developed real-time methods for quantifying gene expression in an appropriate, smoke-exposed epithelium, and comparing that expression to that in precisely-defined microdissected smoke-exposed human lung epithelia. Therefore, the specific aims are: 1) Develop quantitative real-time RT-PCR assays for the evaluation of gene expression of a panel of carcinogen metabolizing enzymes in buccal mucosa, including Ahr, ER, CYP1B1, CYP1A1, GSTM1, GSTM3, GSTP1, GSTT1, NQO1. 2) Compare the gene expression in buccal mucosa to simultaneously sampled plasma assayed for nicotine and cotinine by state-of-the-art LC-MS-MS techniques. 3) Quantitatively compare the expression of carcinogen metabolizing enzyme genes from in situ tobacco-exposed human buccal mucosal cells with that from microdissected human lung bronchial and alveolar epithelial cells also exposed in situ. 4) Measure the variability of this buccal cell carcinogen-metabolizing gene expression across individuals, accounting for expression confounders such as tobacco smoke exposure and dietary constituents, and thereby identify phase I and phase II high or low expressors. The long-term goal is to develop a panel of biomarkers in an easily-accessible, smoke-exposed epithelium that identifies smokers at high-risk for lung cancer, and thereby allows smoking-cessation, chemoprevention and early-disease detection strategies to be focussed on those most at risk.

描述（由申请人提供）： 本建议的具体目标是评估使用基因 在口腔细胞中的表达，以准确反映人肺中的基因表达 当两者都暴露于烟草烟雾时，就像吸烟者一样。的诱导 编码致癌物质代谢酶的基因同时在一个 容易接近的替代组织以及肺表明 吸烟者生物激活基因表达表型的可能性 感应如果这种致癌物质代谢途径确实与肺相关， 癌症易感性，那么高I相和低II相酶 诱导表型可能被假设为赋予肺癌的风险。 肺癌致癌物代谢基因诱导研究的力量 迄今为止，由于受试者数量有限， 适于表达或活性测定的肺组织供应不足。 此外，对这些基因编码序列多态性的研究也 对于肺癌的风险，结果非常复杂。本研究 建议利用新开发的实时方法， 量化适当的烟雾暴露上皮中的基因表达，和 将该表达与精确定义的显微解剖的 暴露于烟雾的人肺上皮细胞。因此，具体目标是：1） 实时荧光定量RT-PCR技术在基因检测中的应用 一组致癌物代谢酶在颊粘膜中的表达， 包括Ahr、ER、CYP 1B 1、CYP 1A 1、GSTM 1、GSTM 3、GSTP 1、GSTT 1、NQO 1。（二） 比较颊粘膜和同时采集的血浆中的基因表达 通过最先进的LC-MS-MS技术测定尼古丁和可替宁。第三章 致癌物代谢酶基因表达的定量比较 从原位烟草暴露的人颊粘膜细胞与从 显微解剖的人肺支气管和肺泡上皮细胞也暴露于 在原地。4)测量口腔细胞的变异性 致癌物代谢基因在个体间的表达，占 表达混杂因素，如烟草烟雾暴露和饮食 成分，从而确定阶段I和阶段II高或低 expressors.长期目标是开发一组生物标志物， 容易接近的，暴露在烟雾中的上皮细胞， 肺癌的高风险，从而允许戒烟， 化学预防和早期疾病检测策略将集中在那些 最危险的