CELL AND MATRIX BIOLOGY OF LIVER CARCINOGENESIS

人们对硬骨鱼用于生物测定测试、检测 环境中的致癌物质，并作为潜在的比较肿瘤学 人类癌症的模型。 然而，基本的问题仍然是发病机制 和生物学特性。 肝脏对 鱼类和高等脊椎动物中的致癌物质。 为了进一步发展鱼类 作为有用的动物模型，我们将分析肝癌发生的各个方面， 在两个有前途的物种，萨尔莫gairdneri和Oryzias latipes。 问题 关于：鱼肝脏的增殖性病变如何符合数据 哺乳动物肝肿瘤;肝细胞的形态学特征 鱼类肿瘤细胞内基质与肝内化疗敏感性的关系 胆管上皮细胞的致癌物质将被解决。 体内偶联 体外研究将集中在鱼类肝细胞群上。 将使用敏感的免疫组织化学探针来确定 对有丝分裂指数、非程序DNA诱导的特异性致癌物 细胞和基质糖蛋白的合成（UDS）和改变， 糖胺聚糖 肿瘤性肝脏病变的过程和命运将 使用改变细胞的酶组织化学和长期 追踪携带肿瘤的鱼。 致癌物质形成特异性 DNA加合物将由超灵敏质谱仪测定 问题研究 我们将寻求将形成与特定的DNA加合物相关联， 肝脏病变的类型。 提出了三个具体目标。 1. 表征病变从正常进展中的形态学事件 日本青鳉鱼肝脏至显性肿瘤发病年龄的函数关系 并确定肝细胞肿瘤的长期潜力， 生长/转移。 2. 在原代培养中分离并维持肝脏 细胞的鳟鱼和开发程序，以分离非肝细胞 人口。 从后者入手，建立长期养殖的鳟鱼胆 上皮细胞 3. 测定肝细胞和其他细胞的能力 类型的鳟鱼肝脏激活遗传毒性致癌物后，在体外 暴露并确定其中的程度和特定细胞类型 致癌物诱导的DNA修复发生。 确定，定性和 定量地，鳟鱼肝脏和细胞中的DNA加合物暴露于各种 致癌物和DNA修复对加合物量的后续影响。 使用 致癌物生物测定，确定DNA加合物的化合物特异性效应 对修复和致癌潜力的影响。

Interest has heightened in use of teleosts for bioassay testing, detection of carcinogens in the environment, and as potential comparative oncology models for human cancer. However, basic questions remain on pathogenesis and biologic properties of fish tumors. The liver is sensitive to carcinogens in fish and higher vertebrates. To further development of fish as useful animal models, we will analyze aspects of hepatic carcinogenesis in two promising species, Salmo gairdneri and Oryzias latipes. Questions about: how proliferative lesions of fish liver conform to data on neoplasms in mammalian liver; morphologic characteristics of liver cells and intracellular matrix in fish neoplasms and sensitivity of intrahepatic biliary epithelial cells to carcinogens will be addressed. Coupled in vivo and in vitro studies will focus on populations of fish liver cells. Sensitive immunohistochemical probes will be used to determine effect of specific carcinogens on mitotic index, induction of unscheduled DNA synthesis (UDS) and alterations of cell and matrix glycoproteins and glycosaminoglycans. Course and fate of neoplastic liver lesions will be studied using enzyme histochemistry of altered cells and long-term follow-up of tumor laden fish. The ability of carcinogens to form specific DNA adducts will be determined by ultrasensitive mass spectrometer studies. We will seek to correlate formation to specific DNA adducts with types of liver lesions. Three specific aims are proposed. 1. Characterize morphologic events in the progression of lesions from normal liver to overt tumor in Japanese medaka as a function of age at initiation and determine long-term potential of liver cell tumors for growth/metastasis. 2. Isolate and maintain, in primary culture, liver cells of trout and develop procedures to separate nonhepatocytic cell populations. From the latter, establish long-term culture of trout biliary epithelial cells. 3. Determine capacity of hepatocytes and other cell types of trout liver to activate genotoxic carcinogens after in vitro exposure and determine extent and specific cell types in which carcinogen-induced DNA repair occurs. Determine, qualitatively and quantitatively, DNA adducts in trout liver and cells exposed to various carcinogens and follow effect of DNA repair on amount of adduct. Using carcinogen bioassay, determine compound specific effects of DNA adduct formation on repair and on carcinogenic potential.