人鼻咽癌DNA对Rat-Ⅰ细胞株的转化活性及其基因表达的研究

为了解EB病毒基因组阳性人鼻咽癌DNA在转染正常细胞后可能发生的生物学作用,作者将人鼻咽癌活检组织的DNA(EB病毒基因组阳性),用磷酸钙方法转染正常Rat-Ⅰ细胞,使其恶转;用软琼脂培养的转化细胞能贴壁生长,并形成克隆;转化细胞易与PHA发生凝集;克隆接种裸鼠形成纤维肉瘤;转化细胞和裸鼠肿瘤组织用抗C_3补体免疫荧光法检测EBNA呈阳性;染色体原位杂交显示在某些细胞中期染色体上有银粒出现。

STUDY OF TRANSFORMATION ACTIVITIES ON RAT-1 AND EXPRESSION OF HUMAN NASOPHARYNGEAL CARCINOMA DNA

Rat-1 cell (rat fibroblast cell line) were transfected with the DNA from biopsy of human nasophar/ngeal carcinoma (NPC, EBV genome positive) by carcium phosphate pre-citate method. The malignant, transformed foci of Rat-1 cell have been cloned, When we use soft agar culture, the transformed cell can grow and form cell colonies. Following passage, the foce were able to adhere to glass wall and grow. These cell were easily agglutinated by PHA. The cloned transformants have been inoculated into nude mice with the formation of fibrosarcoma. The transforming cells and transplanting tumor cells from nude mice were cultured, positive EBV nuclear antigen (EBNA) was detected by anticomplement (C3) immu-nofluencent method. In situ hybridization of the chromosome from transformed cells with EBV-W(YH) (BamH I) probe 3H-labeled showed silaver granules in some metaphase cells.Further study of these transforming cells is helpfull to explain the mechanisms of car-cinogenesis of human NPC.