IN VITRO STUDIES OF HUMAN MESOTHELIAL CELL CARCINOGENESIS

人间皮细胞致癌的体外研究

• 批准号: 3838425
• 负责人: B I GERWIN
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3838425
• 关键词: carcinogenesis; cell line; gene expression; gene mutation; growth factor receptors; human genetic material tag; human tissue; immunocytochemistry; mesothelioma; messenger RNA; molecular oncology; natural gene amplification; neoplasm /cancer genetics; neoplastic cell; platelet derived growth factor; point mutation; retinoblastoma; tissue /cell culture; tumor antigens; tumor suppressor genes

p53 and retinoblstoma tumor suppressor genes have been shown to be altered in many types of human cancer. In addition, in model systems, the combination of a p53 and a ras activating mutation have been required for tumorigenic conversion. We have analyzed a panel of human mesothelioma cell lines for alterations in these genes. In 20 cell lines from 17 individuals, 2 had p53 point mutations and one was a p53 null cell. No Ki-ras activating mutations were detected in any of the 20 cell lines. These studies showed that p53 expression was detectable by immunocytochemistry in normal human pleural mesothelial cells as well as 12 mesothelioma cell lines. The expression level of the human MDM2 gene in mesothelioma cell lines is being investigated since this gene product is frequently found to associate with the p53 protein and is amplified in some tumors in which p53 gene alterations are undetectable. Retinoblastoma gene mRNA expression appeared to be normal and protein studies are ongoing. A ribonuclease protection assay to allow quantitation of mRNA levels for PDGF receptor alpha and beta genes has been developed. This will be applied to human mesothelial cells and mesotheliomas as well as to T antigen immortalized human mesothelial cells overexpressing PDGF-A chain genes and appropriate control cells.

p53 和视网膜母细胞瘤肿瘤抑制基因已被证明 在许多类型的人类癌症中发生改变。此外，在模型系统中， 需要 p53 和 ras 激活突变的组合 致瘤性转化。我们分析了一组人类间皮瘤 改变这些基因的细胞系。在 17 个细胞系的 20 个细胞系中 个体中，2 个具有 p53 点突变，1 个是 p53 无效细胞。不 在 20 个细胞系中的任何一个中都检测到了 Ki-ras 激活突变。 这些研究表明 p53 表达可通过以下方法检测到： 正常人胸膜间皮细胞的免疫细胞化学以及 12 种间皮瘤细胞系。人类MDM2基因的表达水平 由于该基因产物在间皮瘤细胞系中的作用正在被研究 经常被发现与 p53 蛋白相关并在 一些肿瘤中无法检测到 p53 基因改变。 视网膜母细胞瘤基因 mRNA 表达似乎正常，蛋白表达正常 研究正在进行中。 核糖核酸酶保护测定允许 PDGF 受体 α 和 β 基因 mRNA 水平的定量 已开发。这将应用于人类间皮细胞和 间皮瘤以及 T 抗原永生化人间皮瘤 过表达PDGF-A链基因的细胞和适当的对照细胞。