Binding of Benzo(a)pyrene to the Cellular Macromolecules of Mouse Embryonic Fibroblasts in Primary Culture.

국문 초록 (Abstract)

마우스 [NIH (GP)] 태아세포 일차 배양계에 투여된 Benzo(a)pyrene과 세포내 고분자 화합물의 결합량 측정을 시도하였다. $0.5\;{\mu}M\;-8.0\;{\mu}M$의 $^3H$-B(a)P 첨가 배지로 24시간 배양한 후, $^3H$-B(a)P의 세포내외 분포도를 조사한 결과, 가해준 $^3H$-B(a)P의 90% 이상은 배지내에 잔류되어 있었고, 2-10%는 세포 표면에 가볍게 부착되었으며 (alcohol soluble fraction) 0.5%미만은 세포내 고분자 화합물에 결함되었다(alcohol insoluble fraction). B(a)P 24시간 처리군의 세포내 DNA, RNA 단백철 농도는 대조군에 비하여 변화가 없었다. 이때 DNA에 결합되어 냐타나는 $^3H$-B(a)P의 비활성도는 첨가된 B(a)P 농도에 비례하여 증가하다가 $2\;{\mu}M$이상 투여시 $24\;{\mu\mu}moles/mg$으로 포화됨을 확인하였다. 단백질과의 결합량은 B(a)p $2\;{\mu}m$ 까지는 농도에 비례증가하였으며, $8\;{\mu}m$까지 B(a)P와 단백질 결합이 계속 증가하는 것을 확인 하였다(r=0.99).

다국어 초록 (Multilingual Abstract)

We describe here the binding of benzo(a)pyrene to the cellular macromolecules of mouse embryonic fibroblasts in primary culture. On the 3rd day of the culture, DME medium containing 10% FCS was removed and the cells were incubated with fresh medium containing $1\;{\mu}M$, $2\;{\mu}M$ and $4\;{\mu}M$ of benzo(a)pyrene for 24 hours. Cell growh was inhibited more than 20% after the first 48 hours and more than 50% after 96 hours. B(a)p cytotoxicity was not so severe with the 24 hour treatment. $^3H$-B(a)P was applied to the cell monolayers and more than 90% remained in the culture medium after a 24 hr incubation and about 2 to 10% was loosely bound to the cell surface. The $^3H$-B(a)P tightly bound to the cellular macromolecules was less than 0.5%. The contents of cellular DNA, RNA and protein were not changed with the treatment of 2 to $8\;{\mu}M$ of B(a)P for 24 hours. Specific binding of $^3H$-B(a)P to DNA was linearly increased depending on the concentration of B(a)p lower than $2\;{\mu}M$, and it was saturated at $24\;{\mu\mu}moles/mg$ with higher than $2\;{\mu}m$ of B(a)p. Specific activity of $^3H$-B(a)P-protein-protein adduct was also lineraly increased with $2\;{\mu}m$ of B(a)p, but it was not saturated up to $8\;{\mu}M$ of B(a)p.