Kampinga H H, Mullenders L H, Konings A W
Department of Radiopathology, State University of Groningen, The Netherlands.
Int J Radiat Biol Relat Stud Phys Chem Med. 1988 Feb;53(2):291-300. doi: 10.1080/09553008814550641.
Nuclear matrices of heated and non-heated HeLa S3 cells were isolated and average DNA loop-sizes were compared. Heat treatment (30 min at 45 degrees C) resulted in an ultimate survival level of the cells of about 10 per cent. The loop-size determinations were done on nuclear material isolated from the cells directly after heat treatment. In the nuclear matrices isolated from the heated cells about 1.8 times more protein was bound as compared to the matrices from control cells. Enzymatic analysis using DNase I digestion, followed by centrifugation on neutral sucrose gradients, was performed. Also, halo visualization was combined with autoradiography. Both methods revealed no gross alterations in DNA loop-sizes. The possible function of DNA loop organization in the effect of hyperthermic interference with DNA-related processes is discussed.
分离了经加热和未经加热的海拉S3细胞的核基质，并比较了平均DNA环大小。热处理（45℃ 30分钟）导致细胞的最终存活水平约为10%。环大小的测定是在热处理后直接从细胞中分离出的核物质上进行的。与对照细胞的核基质相比，从受热细胞中分离出的核基质结合的蛋白质多出约1.8倍。使用DNA酶I消化，然后在中性蔗糖梯度上离心进行酶分析。此外，将晕圈可视化与放射自显影相结合。两种方法均未显示DNA环大小有明显变化。讨论了DNA环组织在热干扰与DNA相关过程中的可能作用。
