CHEMICAL TRANSFORMATION OF HUMAN LYMPHOBLASTOID CELL LINES

这项计划的目的是描述化学物质的性质。 诱导从良性过度增殖向恶性转化 爱泼斯坦-巴尔病毒(EBV)使人淋巴细胞永生。通过以下方式治疗 N-乙酰氧基-2-乙酰氨基荧烯(N-OAc-AAF)是一种有效的移码诱变剂， EB病毒永生化淋巴细胞向高级淋巴细胞的诱导转化 “免疫母细胞性淋巴瘤”注射到裸鼠体内，而 注射未经处理的原始细胞则不起作用。肿瘤细胞是 所有的B细胞血统。从化学上提取的高分子量DNA 转化的EBV永生化人脐带血淋巴细胞 PSV2Neo共转染NIH3T3细胞。在2周后选择 250微克/mlG418，克隆出10个病灶并扩增。这些焦点 软琼脂生长呈阳性，并在2小时内出现纤维肉瘤 在无菌小鼠体内存活数周。对照组NIH3T3细胞不能 非锚定生长或肿瘤形成。后来的南区 分析表明，原代转染的3T3细胞含有 大约95kbp的人类DNA，通过与 人类重复序列，蓝色8。各种癌基因病灶的筛选 结果表明，c-raf-1基因的3‘端存在于细胞内。 这是关于c-raf-1化学激活的第一个报告。 人类细胞系中的癌基因。

The object of this project is to characterize the nature of chemically induced transition from a benign hyperproliferative to a malignant state in Epstein-Barr virus (EBV) immortalized human lymphocytes. Treatment with N-acetoxy-2-acetyl-aminofluorene (N-OAc-AAF), a potent frameshift mutagen, induced conversion of the EBV immortalized lymphocytes into high grade "immunoblastic lymphomas" upon injection into athymic mice, whereas injection of the untreated, original cells did not. The tumor cells were all of the B-cell lineage. High molecular weight DNA from the chemically transformed EBV immortalized human cord blood lymphocytes was co-transfected with pSV2Neo into NIH 3T3 cells. After 2 weeks selection in 250 micrograms/ml G418, 10 foci were cloned out and expanded. These foci were positive for soft agar growth and gave rise to fibrosarcomas within 2 weeks in athymic mice. The control NIH 3T3 cells were incapable of anchorage-independent growth or tumor formation. Subsequent Southern analysis revealed that the primary transfected 3T3 cells contained approximately 95 kbp of human DNA as indicated by hybridization to the human repeat sequence, Blue 8. Screening of the foci for various oncogenes revealed that the 3' end of c-raf-1 was present in the transfected cells. This constitutes the first report of chemical activation of the c-raf-1 oncogene in human cell lines.