Mutagenicity of tobacco smoke in human cell co-cultures

人类细胞共培养物中烟草烟雾的致突变性

• 批准号: 7267989
• 负责人: JOSEPH B GUTTENPLAN
• 金额: $ 16.98万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-18 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7267989
• 关键词: Abbreviations; Applications Grants; Area; Benzo(a)pyrene; Biological Assay; Biological Models; Cancer Etiology; Cancer Model; Carcinogens; Cell Line; Cells; Cigarette; Coculture Techniques; Cytochrome P450; Detection; Dose; Embryo; End Point; Environmental Carcinogens; Epithelial; Epithelial Cells; Event; Gene Expression; Genetic; Goals; Green Fluorescent Proteins; Human; Incidence; Individual; Link; Malignant Neoplasms; Marketing; Metabolic Activation; Metabolism; Modeling; Mutagenesis; Mutagens; Mutation; N' -nitrosonornicotine; NIH Program Announcements; Numbers; Oral; Oral cavity; Oral mucous membrane structure; Organ; Pilot Projects; Rattus; Relative (related person); Reporter; Reporter Genes; Research; Reverse Transcriptase Polymerase Chain Reaction; Risk; Shuttle Vectors; Site; Site-Directed Mutagenesis; Smoke; Smoking Behavior; System; Techniques; Testing; Tobacco; Tobacco smoke; Tobacco smoking; Tobacco-Associated Carcinogen; Tongue; Toxic effect; Transgenic Organisms; Urination; Variant; Western Blotting; base; carcinogenesis; carcinogenicity; experience; in vitro Model; in vivo; interest; malignant mouth neoplasm; monolayer; mutant; novel; tobacco exposure; tobacco tar

DESCRIPTION (provided by applicant): Tobacco smoking is a major cause of cancers at many sites, particularly the aerodigestive tract. For many years, filter and reduced tar cigarettes have been available, but have not resulted in reduced incidences of smoking-related cancers, because of compensatory smoking behavior. Recently, newer versions of "reduced-risk" cigarettes have been marketed, but their relative carcinogenicities are unknown. Cancer arises through multistage carcinogenesis, which involves the accumulation of genetic damage (such as mutations), over years, until a cell converts to cancer. Current assays for potential carcinogenesis in humans all have certain deficiencies. Few models employ normal human cells and mutagenesis is generally not an endpoint. To better understand risk from potential carcinogens, new model systems that can evaluate such risk are needed. In preliminary studies, we showed that in monolayer co-culture of normal human oral epithelial cells (NOE's) with a lac I rat reporter cell line (BB cells - that contain a lambda-based shuttle vector for detecting mutagenesis), increased mutagenesis induced by benzo(a)pyrene (BaP), relative to BB cells alone. This increased mutagenesis is most likely due to activation of BaP by the NOE's. We hypothesize that co-culturing NOE's with lac I reporter cells will allow detection of mutagenesis that is causally linked to metabolic activation of carcinogens by the normal human oral cells. To validate the model, mutagenesis induced by the two tobacco carcinogens (BaP and nitrosonornicotine) will be investigated. In Aim 1 we will establish and characterize a co- culture model composed of NOE's co-cultured with BB reporter cells. In Aim 2, this model will be used: 1) to compare mutagenesis induced by tobacco smoke condensate (TSC) from conventional cigarettes and recently-introduced "low-risk" cigarettes and 2) to determine inter-individual variation in the metabolism of these TSC's. Since human cells from a variety of organ sites can be grown in culture, the model has potential to become a flexible broad-based system to evaluate carcinogenicity risk of tobacco products in a number of human organs. A third aim will be a pilot study on the levels of cytochrome P- 450's in NOE's with and without the presence of TSC's, using gene expression arrays and confirmation by RT-PCR and western blotting. As NOE's are the very cells that are destined to develop into oral cancers, the model fills a void with the potential of being an organ specific technique to quantify mutagenesis, a major step in carcinogenesis. Clearly, it is preferable to determine their potential risk prospectively, rather than retrospectively.

描述（由申请人提供）：吸烟是许多部位癌症的主要原因，特别是呼吸消化道。多年来，过滤嘴香烟和低焦油香烟一直存在，但由于补偿性吸烟行为，并没有导致吸烟相关癌症的发病率降低。最近，新版本的“降低风险”香烟已经上市，但其相对致癌性尚不清楚。癌症是通过多阶段致癌作用产生的，这涉及遗传损伤（如突变）的积累，多年来，直到细胞转化为癌症。目前对人类潜在致癌作用的测定都有一定的缺陷。很少有模型采用正常的人类细胞，诱变通常不是终点。为了更好地了解潜在致癌物的风险，需要能够评估这种风险的新模型系统。在初步研究中，我们表明，在正常人口腔上皮细胞（NOE）与lac I大鼠报告细胞系（BB细胞-含有用于检测诱变的基于DNA的穿梭载体）的单层共培养中，相对于单独的BB细胞，苯并（a）芘（BaP）诱导的诱变增加。这种增加的诱变很可能是由于NOE激活BaP所致。我们假设NOE与lac I报告细胞共培养将允许检测与正常人口腔细胞的致癌物代谢活化有因果关系的诱变。为了验证该模型，将研究两种烟草致癌物（BaP和亚硝基去甲烟碱）诱导的诱变。在目标1中，我们将建立和表征由NOE与BB报告细胞共培养组成的共培养模型。在目标2中，该模型将用于：1）比较来自常规香烟和最近引入的“低风险”香烟的烟草烟雾冷凝物（TSC）诱导的诱变，和2）确定这些TSC代谢的个体间差异。由于来自各种器官部位的人体细胞可以在培养物中生长，该模型有可能成为一个灵活的基础广泛的系统，以评估烟草产品在许多人体器官中的致癌风险。第三个目标是使用基因表达阵列并通过RT-PCR和蛋白质印迹法进行确认，对存在和不存在TSC的NOE中细胞色素P- 450的水平进行初步研究。由于NOE是注定要发展成口腔癌的细胞，该模型填补了一个空白，具有作为器官特异性技术来量化诱变的潜力，诱变是致癌作用的主要步骤。显然，最好是前瞻性地而不是回顾性地确定其潜在风险。

项目成果

Concentration dependent effects of tobacco particulates from different types of cigarettes on expression of drug metabolizing proteins, and benzo(a)pyrene metabolism in primary normal human oral epithelial cells.

来自不同类型的香烟的烟草颗粒物对药物代谢蛋白表达的浓度依赖性作用，以及原发性正常人口腔上皮细胞中苯并（A）pyrene代谢的浓度。

• DOI: 10.1016/j.fct.2011.06.037
• 发表时间: 2011-09
• 期刊: FOOD AND CHEMICAL TOXICOLOGY
• 影响因子: 4.3
• 作者: Sacks, Peter G.;Zhao, Zhong-Lin;Kosinska, Wieslawa;Fleisher, Kenneth E.;Gordon, Terry;Guttenplan, Joseph B.
• 通讯作者: Guttenplan, Joseph B.