Mironov N M
Biull Eksp Biol Med. 1987 Oct;104(10):486-8.
The confluent culture of hamster embryo cells was incubated with benzo(a)pyrene for 24 hours. Then the medium was replaced by maximal lacking both the serum and benzo(a)pyrene. The process of DNA repair was observed in four nuclear fractions according to two indexes: the disappearance of metabolites of benzo(a)pyrene covalently bound to DNA and the incorporation of 3H-thymidine to DNA in the period from I min to 72 hours. Hydroxyurea at the concentration of 5 mM was added 2-19 hours before 3H-thymidine. The highest concentration of benzo(a)pyrene metabolites was found in the DNA of nuclear matrix fraction throughout all the experiment. The initial concentration of 3H-thymidine right after its addition into the cell culture medium was the highest in DNA of nuclear matrix fraction and the lowest in DNA fraction soluble in the buffer with low ionic strength. Later on, the concentration of 3H-thymidine was decreased in matrix-bound fractions and increased in other fractions up to the total DNA level. The results suggest that the repair process requires joining of benzo(a)pyrene damaged DNA region to the nuclear matrix with the following reverse transition into the fraction where the fragment was initially located.
将仓鼠胚胎细胞的汇合培养物与苯并(a)芘一起孵育24小时。然后用既不含血清也不含苯并(a)芘的培养基进行更换。根据两个指标在四个核组分中观察DNA修复过程:与DNA共价结合的苯并(a)芘代谢产物的消失以及在1分钟至72小时期间3H-胸腺嘧啶核苷掺入DNA的情况。在加入3H-胸腺嘧啶核苷之前2至19小时加入浓度为5 mM的羟基脲。在整个实验过程中,在核基质组分的DNA中发现苯并(a)芘代谢产物的浓度最高。刚加入细胞培养基后3H-胸腺嘧啶核苷的初始浓度在核基质组分的DNA中最高,而在低离子强度缓冲液可溶的DNA组分中最低。随后,与基质结合的组分中3H-胸腺嘧啶核苷的浓度降低,而其他组分中的浓度升高直至达到总DNA水平。结果表明,修复过程需要将苯并(a)芘损伤的DNA区域连接到核基质,随后反向转变为片段最初所在的组分。