喉癌中的癌基因：转化细胞DNA的连续传代

Oncogenes in laryngeal cancer: serial passage of transformed cellular DNA.

作者信息

Friedman W H, Rosenblum B N, Loewenstein P, Thornton H, Katsantonis G, Green M

出版信息

Otolaryngol Head Neck Surg. 1985 Jun;93(3):346-50. doi: 10.1177/019459988509300311.

DOI:10.1177/019459988509300311

Abstract

DNA originally extracted from squamous cell cancer of the larynx has been serially passaged through transformed populations of NIH/3T3 mouse fibroblasts. The transformed foci were then harvested, cloned to volume, and incubated with a fresh population of NIH/3T3 cells in a second passage. Transforming efficiencies were enhanced by serial passage. In addition, Southern Blot analysis of the transformed foci revealed hybridization between transformant DNA and human probe DNA from the Alu family of conserved human DNA sequences. In the first passage this hybridization took the form of diffuse homology throughout the entire molecular weight distribution. The second-passage DNA showed "narrow bands" indicating the possibility that an oncogene has been identified in laryngeal cancer and that serial passage has eliminated contaminating human sequences. Repetitive transfection in third- and fourth-passage studies is now being completed.

摘要

最初从喉鳞状细胞癌中提取的DNA已连续通过经转化的NIH/3T3小鼠成纤维细胞群体传代培养。然后收获转化灶，克隆扩增，并在第二轮传代培养中与新的NIH/3T3细胞群体一起孵育。连续传代提高了转化效率。此外，对转化灶的Southern印迹分析显示，转化体DNA与来自人类保守DNA序列Alu家族的人类探针DNA之间发生了杂交。在第一轮传代中，这种杂交表现为在整个分子量分布范围内的弥漫性同源性。第二轮传代的DNA显示出“窄带”，这表明有可能在喉癌中鉴定出一种癌基因，并且连续传代已消除了污染的人类序列。目前正在完成第三轮和第四轮传代研究中的重复转染。