ONCOGENE EXPRESSION IN HUMAN CARCINOMAS

人类癌症中癌基因的表达

• 批准号: 3939338
• 负责人: P H HAND
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 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/3939338
• 关键词: Escherichia; breast neoplasms; colon disorder; complementary DNA; fibroma; gastrointestinal epithelium; gene expression; gene mutation; genetic manipulation; immunochemistry; immunoperoxidase; mammary disorder; monoclonal antibody; neoplasm /cancer genetics; oncogenes; oncoproteins; radioimmunoassay; viral carcinogenesis

Several distinct and high-conserved genes comprise the ras gene family of rodents and humans, i.e., rodent Harvey and Kirsten, and human Harvey, Kirsten and neuroblastoma. Transformation, either by a point-mutation resulting in a change in one amino acid of the 21 kDa ras gene product (p21), or by increased expression of ras p21, has been demonstrated to be mediated by members of this gene family. We have reported the development of direct binding liquid competition radioimmunoassays for the detection and quantitation of the ras oncogene and proto-oncogene products. Using these radioimmunoassays and ras p21 purified from Escherichia coli containing the full-length T24 mutant human Harvey ras gene protein product as a standard, we have defined the actual amount of ras p21 per mu g of total cellular protein, or per cell, in various ras transformed and "normal" mammalian cell lines. Absolute levels of Ha-ras p21 have also been determined in human breast and colon carcinomas, benign lesions, and/or their respective normal tissues using the radioimmunoassays. Enhanced Ha-ras expression was documented in 66% of breast and 100% of colon carcinomas as compared with their normal counterparts, with levels in breast carcinomas ranging from 18.4 to 51.7 pg ras p21 per mu g protein. Some dysplastic lesions of the breast and colon also contained elevated Ha-ras p21. Relative levels of Ha-ras p21 expression, detected by competition RIA, correlated with percent Ha-ras p21 positive cells as determined by immunohistochemical assays. Using liquid competition RIA and immunohistochemical assays, it has been shown that levels of ras p21 expression did not always correlate between primary and metastatic colon lesions of the same patient. The use of the quantitative RIA and semiquantitative immunohistochemical assays, in concert with cDNA probes for identification of specific ras pointmutated oncogenes or protooncogenes, may now provide the means for definitive quantitative analyses of ras p21 in human carcinomas and benign lesions.

Ras基因由几个不同的高度保守的基因组成。 啮齿动物和人类的家族，即啮齿动物哈维和柯尔斯滕， 以及人类的哈维、柯尔斯滕和神经母细胞瘤。转型， 要么是通过点突变导致一种氨基酸的变化 21 kDa ras基因产物(P21)，或通过增加表达 Ras p21，已被证明是由 这个基因家族。我们已经报告了Direct的发展情况 结合型液体竞争放射免疫分析法检测 Ras癌基因和原癌基因的定量检测 产品。用这些放射免疫分析和纯化的ras p21 从含有全长T24突变体的大肠杆菌中分离到 以人哈维ras基因蛋白产品为标准，我们有 定义了每微克细胞总数的ras p21的实际数量 蛋白质，或每个细胞，在不同的RAS中转化和“正常” 哺乳动物细胞系。Ha-ras p21的绝对水平也 在人类乳腺癌和结肠癌中被检测出是良性的 病变和/或其各自的正常组织使用 放射免疫分析。记录了增强的Ha-ras表达 在66%的乳腺癌和100%的结肠癌中 它们的正常对应物，在乳腺癌中的水平 每单位蛋白18.4至51.7pg ras p21。一些人 乳腺和结肠的异常增生性病变也含有隆起 Ha-ras p21。Ha-ras p21表达的相对水平 竞争RIA与Ha-ras p21阳性百分率相关 免疫组织化学方法检测细胞数。使用液体 竞争RIA和免疫组织化学分析，它已经 表明ras p21的表达水平并不总是相关的 原发灶和转移灶之间的关系 有耐心的。定量放射免疫法和半定量放射免疫法的应用 免疫组织化学检测，结合cDNAs探针 特异性ras点突变癌基因的鉴定 原癌基因，现在可能提供最终的手段 人癌和良性病变中ras-p21的定量研究 损伤。