SPRL--A MOLECULAR MARKER FOR BRONCHIAL PRENEOPLASIA

SPRL--支气管癌前病变的分子标记

• 批准号: 2443219
• 负责人: DERICK LAU
• 金额: $ 7.66万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2443219
• 关键词: DNA footprinting; biomarker; bronchus neoplasm; cell transformation; cellular oncology; gel mobility shift assay; gene expression; genetic regulatory element; human tissue; immunocytochemistry; in situ hybridization; metastasis; molecular cloning; nuclear runoff assay; oncoproteins; preneoplastic state; proline; respiratory epithelium; squamous cell carcinoma; tissue /cell culture; transcription factor

DESCRIPTION (Applicant's Description): This program is designed to establish the applicant as an independent investigator of biomarker research in bronchial carcinogenesis, and in early detection and chemoprevention of cancers of the upper aerodigestive tract. The applicant has previous research experience in molecular and cellular pharmacology and has recently become an established medical oncologist specializing in the treatment of lung cancer. The sponsor is an accomplished pulmonary physiologist and molecular biologist in the field of development and differentiation of bronchial epithelium, and in the area of regulation of biomarkers of the bronchial epithelium in response to environmental toxins such as ozone and tobacco smoke. The plans of the sponsor are to provide supervision to the applicant to acquire expertise in molecular techniques, in the design and execution of experiments to elucidate the mechanisms of regulation of a molecular marker, spr1, associated with bronchial metaplasia, and in the application of this biomarker for early detection of lung cancer. The sponsor has cloned a small proline-rich protein (spr-1) which is overexpressed in tracheobronchial epithelium undergoing metaplastic change from a normal mucociliary phenotype to a squamous appearance. The spr1 expression is down-regulated by vitamin A and up-regulated by tumor promoters. Therefore, it appears that spr1 is a potential early biomarker for preneoplastic transformation of the bronchial epithelium. In their laboratory, there is a series of human bronchial epithelial cells displaying varied degree of spr1 expression and tumorigenicity potential. This proposal is, thus, intended to use this cell-culture model to elucidate the mechanism of regulation of spr1 expression in the multistep carcinogenesis of the bronchial epithelium, and to use archival specimens to map spr1 expression in the field of cancerization of lung tissues from patients with lung cancer. The first aim is to test the hypothesis that there is an increasing expression of spr1 in the field of cancerization extending from a focus of bronchial squamous carcinoma to the surrounding dysplastic, metaplastic and normal bronchial epithelia as determined by immunohistochemical staining and in situ hybridization of archival human squamous lung carcinoma. The second aim is to use a human tracheobronchial cell model to test the hypothesis that the cell type-specific expression of spr1 is regulated at the level of transcription by using a nuclear run-on transcriptional assay and a transfection study with chimeric constructs of the spr1 promoter region attached to a reporter gene. The third aim is to identify the cis-elements and the transcriptional factors involved in the regulation of spr1 gene by employing the techniques of DNA-binding mobility shift gel assay, DNA footprinting and expression cloning. These studies are to set the stage for using spr1 as a biomarker for identifying the early steps of multistep carcinogenesis, for early detection of lung cancer, and for testing the efficacy and mechanisms of chemopreventive agents in reversing malignant transformation of the bronchial epithelium.

描述（申请人的描述）：此程序旨在 将申请人建立为生物标志物研究的独立研究者 在支气管癌发生，以及早期检测和化学预防中 上层机修区的癌症。 申请人以前有 分子和细胞药理学方面的研究经验，最近有 成为专门治疗的既定医学肿瘤学家 肺癌。 赞助商是一位经验丰富的肺部生理学家和 分子生物学家在发展和分化领域 支气管上皮，以及在生物标志物的调节区域 响应环境毒素（例如臭氧和）的支气管上皮 烟草烟。 赞助商的计划是为 申请人获得分子技术，设计和 执行实验以阐明调节机制 分子标记物，Spr1，与支气管化生相关，在 该生物标志物在肺癌的早期检测中的应用。 赞助商克隆了一个富含脯氨酸的小蛋白（SPR-1） 过表达的气管支气管上皮，经历了化学变化 从正常的粘膜纤毛表型到鳞状外观。 SPR1 表达被维生素A下调并被肿瘤上调 发起人。 因此，看来SPR1是潜在的早期生物标志物 用于支气管上皮的前塑性转化。 在他们的 实验室，有一系列显示的人支气管上皮细胞显示 SPR1表达和肿瘤性潜力的多样性。 这 因此，提案旨在使用此细胞培养模型来阐明 多步癌发生中SPR1表达调控的机理 支气管上皮，并使用档案标本映射SPR1 患者的肺组织癌化领域的表达 肺癌。 第一个目的是检验越来越多的假设 SPR1在癌化领域的表达从一个重点延伸 支气管鳞状癌对周围的发育不良，变质和 通过免疫组织化学染色确定的正常支气管上皮和 档案库人鳞状癌的原位杂交。 第二个 目的是使用人类气管支出细胞模型来检验假设 SPR1的细胞类型特异性表达受到调节 使用核跑步转录测定和A转录 使用SPR1启动子区域的嵌合构建体的转染研究 附着在记者基因上。 第三个目的是确定顺式元素 以及通过调节SPR1基因的转录因子 采用DNA结合迁移率凝胶测定的技术，DNA 足迹和表达克隆。 这些研究是为使用SPR1作为生物标志物的阶段 确定多步癌发生的早期步骤，以供早期检测 肺癌，以及测试的功效和机制 化学预防药物逆转恶性转化 支气管上皮。