MOLECULAR CLONING OF TUMOR ASSOCIATED ANTIGENS

单抗技术的使用使得 人类癌症的鉴定和部分特征- 猪瘟的相关抗原和分化抗原 乳腺和结肠上皮。单抗提供了强大的工具 在人类癌症诊断和治疗中的应用。他们 已被用于筛查和诊断目的 在血清检测中检测肿瘤相关抗原，以及 原发癌和转移癌的放射定位 原位损伤。使用放射性标记的单抗免疫球蛋白 和碎片，一种强大的治疗方案正在开发中。 这个项目的总体目标是从分子上克隆和 确定编码肿瘤相关抗原的基因 TAG-72和CEA。我们最近启动了对 TAG-72的克隆。这些研究包括：1)建造 LS174基因文库在几种原核生物和 真核表达载体。2)LS174的共转染 携带药物选择标记物的质粒进入肿瘤DNA 哺乳动物细胞。3)寡聚探针的构建 TAG-72的多肽序列用作杂交探针。 最近启动了更多的研究，以研究 CEA在结肠和乳腺肿瘤细胞系中的表达调控 干扰素诱导后。几个DNA探针已经被 用于Poly A精选的Northern印迹分析 从这些细胞系中提取的mRNA。

The use of monoclonal antibody technology has allowed the identification and partial characterization of human carcinoma- associated antigens as well as differentiation antigens of mammary and colon epithelium. MAbs provide powerful tools for use in the diagnosis and management of human carcinomas. They have been used for screening and diagnostic purposes for the detection of tumor associated antigens in blood serum assays, and for the radiolocalization of primary and metastatic carcinoma lesions in-situ. Using radiolabeled monoclonal immunoglobulins and fragments, a powerful therapy regime is being developed. The overall goal of this project is to molecularly clone and identify the genes that encode the tumor associated antigens TAG-72 and CEA. We have recently initiated studies for the cloning of TAG-72. These studies include: 1) the construction of cDNA libraries from LS174 mRNA in several prokaryotic and eukaryotic expression vectors. 2) the cotransfection of LS174 tumor DNA with plasmids carrying drug selectable markers into mammalian cells. 3) the construction of oligomeric probes to the peptide sequence of TAG-72 for use as hybridization probes. Additional studies have recently been initiated to study the regulation of CEA expression in colon and breast tumor cell lines after interferon induction. Several DNA probes have been developed to use in Northern blot analysis of poly A selected mRNA from these cell lines.