Cone R D, Grodzicker T, Jaramillo M
Cold Spring Harbor Laboratory, New York 11724.
Mol Cell Biol. 1988 Mar;8(3):1036-44. doi: 10.1128/mcb.8.3.1036-1044.1988.
An epithelial cell-transforming virus could be of great use, both in the culture of epithelial cell lines and in the study of carcinogenesis. Since the adenoviral E1A gene has been shown to partially transform some epithelial cells from primary rat cell cultures, we constructed retrovirus vectors containing either the 12S or 13S E1A cDNA sequences to facilitate the transfer of these genes into a variety of primary cell types. The 12S E1A virus induced proliferation and immortalization of epithelial cells in rat kidney, liver, heart, pancreas, and thyroid primary cultures. In the two cases tested, heart and liver cultures, E1A-immortalized cells were nontumorigenic, but could be completely transformed by subsequent introduction of the ras oncogene. To our surprise, the 13S virus had a greatly reduced immortalization potential. We discuss these data in light of the model of Spindler et al. (K. R. Spindler, C. Y. Eng, and A.-J. Berk, J. Virol. 53:742-750, 1985), in which the 12S E1A protein is required for the complete induction of the cellular DNA replication machinery in the quiescent human epithelial cells in which adenoviruses normally replicate.
一种上皮细胞转化病毒在培养上皮细胞系和研究致癌作用方面都可能有很大用途。由于腺病毒E1A基因已被证明能使原代大鼠细胞培养中的一些上皮细胞部分转化,我们构建了含有12S或13S E1A cDNA序列的逆转录病毒载体,以便将这些基因转移到多种原代细胞类型中。12S E1A病毒在大鼠肾脏、肝脏、心脏、胰腺和甲状腺原代培养物中诱导上皮细胞增殖并使其永生化。在测试的两个案例中,即心脏和肝脏培养物,E1A永生化细胞无致瘤性,但随后引入ras癌基因可使其完全转化。令我们惊讶的是,13S病毒的永生化潜力大大降低。我们根据Spindler等人的模型(K. R. Spindler、C. Y. Eng和A.-J. Berk,《病毒学杂志》53:742 - 750,1985年)来讨论这些数据,在该模型中,12S E1A蛋白是在腺病毒正常复制的静止人上皮细胞中完全诱导细胞DNA复制机制所必需的。