热疗对HeLa S3细胞DNA环大小的影响。

H H Kampinga, L H Mullenders, A W Konings
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引用次数: 7

摘要

分离加热和未加热的HeLa S3细胞的核基质,比较平均DNA环的大小。热处理(在45℃下30分钟)导致细胞的最终存活水平约为10%。循环大小的测定是在热处理后直接从细胞中分离的核材料上进行的。在从加热细胞分离的核基质中,与对照细胞的基质相比,结合的蛋白质约为1.8倍。使用DNase I酶切进行酶分析,然后在中性蔗糖梯度上离心。同时,光晕可视化与放射自显影相结合。两种方法均未发现DNA环大小的明显变化。讨论了DNA环组织在高温干扰DNA相关过程中的可能作用。
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Effect of hyperthermia on DNA loop-size in HeLa S3 cells.

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.

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