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目的研究苯并(a)芘[B(a)P]对体外培养的人胚肺成纤维细胞(HELF)中P53、P21蛋白量的影响及芳烃受体(AhR)途径的作用。方法在有/无芳烃受体拮抗剂α-萘黄酮预孵下,分别在-/+体外代谢活化系统(S9)的条件下,以B(a)P染毒HELF细胞,用蛋白印迹(Westernblot)方法检测细胞P53、P21蛋白量的改变。结果10,20,50μmol/LB(a)P在-/+S9时染毒2h,以及在-S9时染毒24h引起细胞P53、P21蛋白量升高。B(a)P在-/+S9染毒2h时所引起的P53蛋白量升高可分别被2.5和5.0μmol/Lα-萘黄酮(α-NF)预先孵育逆转,所引起的P21蛋白量的升高可分别被10.0μ和2.5～10.0μmol/Lα-NF预孵逆转。B(a)P在-S9染毒24h时所引起的P53蛋白量升高可被2.5和5.0μmol/Lα-NF预孵逆转,所引起的P21蛋白量升高可被2.5和5.0μmol/Lα-NF预孵逆转。结论P53、P21蛋白在HELF细胞应对B(a)P引起DNA损伤的反应机制中起关键性作用,芳烃受体途径在其中存在一定作用。
Objective To investigate the effect of benzo (a) pyrene [B (a) P] on the protein levels of P53 and P21 in human embryo lung fibroblasts (HELF) cultured in vitro and the effect of aromatase receptor (AhR) pathway. Methods HELF cells were exposed to B (a) P preincubated with α-naphthalene flavone with or without aromatics receptor antagonist in the presence of - / + in vitro metabolic activation system (S9) ) Method was used to detect the changes of P53 and P21 protein in cells. Results 10, 20, 50μmol / L (a) P exposure to - / + S9 for 2h, and exposure to - S9 for 24h resulted in increased cellular P53, P21 protein levels. The increase of P53 protein induced by B (a) P at + / - S9h could be reversed by preincubation with 2.5 and 5.0μmol / L α-NF, respectively. The amount of P21 protein Elevated by 10.0μ and 2.5 ~ 10.0μmol / L α-NF pre-incubation reversed. The increase of P53 protein induced by B (a) P at -S9 for 24h could be reversed by preincubation with 2.5 and 5.0μmol / Lα-NF, and the increase of P21 protein was induced by 2.5 and 5.0μmol / Lα -NF pre-incubation reversed. Conclusions P53 and P21 proteins play key roles in response to DNA damage induced by B (a) P in HELF cells. Aromatase receptor pathway may play a role in this process.