Krystosek A, Puck T T
Eleanor Roosevelt Institute for Cancer Research, Denver, CO 80206.
Proc Natl Acad Sci U S A. 1990 Sep;87(17):6560-4. doi: 10.1073/pnas.87.17.6560.
The malignant CHO-K1 cell is reverse-transformed by cAMP, regaining the phenotype of a normal fibroblast. During this reaction, much of its DNA re-acquires sensitivity to hydrolysis by DNase I in a way characteristic of the normal fibroblast. Exposed DNA forms a rim about the nucleus in both the normal and reverse-transformed cell but not in the malignant CHO-K1. Reacquisition of the nuclear rim requires an organized cytoskeleton. Sequestered DNA forms families of different degrees of sequestration. In accordance with previous theoretical developments it is proposed that (i) genes specific to a given differentiation state are stored in the nuclear rim, whereas genes specific to other states are sequestered within the nucleus; (ii) only exposed genes are active, and their activity is modulated by regulatory molecules in the fluid medium; (iii) exposure and sequestration are regulated by cytoskeletal and nuclear protein structures; (iv) in at least several types of cancer the regulatory defect lies in the genome exposure process so that the specific DNA sequences and their associated growth regulatory loci have been transferred from the exposed to the sequestered condition with consequent loss of the nuclear rim of exposed DNA. The methodology described should be generally applicable to examining the accessibility state of subsets of DNA during various physiological modulations of cell function.
恶性CHO - K1细胞被环磷酸腺苷(cAMP)逆转转化,恢复为正常成纤维细胞的表型。在这个反应过程中,其大部分DNA重新获得了对脱氧核糖核酸酶I(DNase I)水解的敏感性,这是正常成纤维细胞特有的方式。在正常细胞和逆转转化细胞中,暴露的DNA在细胞核周围形成一个边缘,但在恶性CHO - K1细胞中则没有。重新获得核边缘需要一个有组织的细胞骨架。被隔离的DNA形成了不同隔离程度的家族。根据先前的理论发展,提出以下观点:(i)特定分化状态的基因存储在核边缘,而其他状态的基因则被隔离在细胞核内;(ii)只有暴露的基因是活跃的,其活性由流体介质中的调节分子调节;(iii)暴露和隔离由细胞骨架和核蛋白结构调节;(iv)在至少几种类型的癌症中,调节缺陷在于基因组暴露过程,使得特定的DNA序列及其相关的生长调节位点从暴露状态转移到了隔离状态,从而导致暴露DNA的核边缘丧失。所描述的方法通常应适用于在细胞功能的各种生理调节过程中检查DNA子集的可及性状态。