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目的探讨苯并(a)芘对人子宫内膜雌激素代谢相关基因作用的分子机制。方法采用人子宫内膜癌细胞株进行体外实验,研究苯并(a)芘对细胞色素P450酶的作用,以0.016、0.8、0.4、2.0、10μmol/LBP作用RL95-2细胞12h,2μmol/LBP作用RL95-2细胞6、12、24、36、48h,细胞经收集、裂解后,用Westernblot分析方法检测总细胞提取液中CYP1A1、CYP1B1和CYP3A4蛋白表达,并用表达了CYP450酶的supersomes作为阳性对照来检测方法的可靠性。结果不同剂量BP作用于RL95-2细胞12h后,CYP1A1蛋白表达明显增加,2μmol/LBP作用于RL95-2细胞不同时间后,CYP1A1随作用时间延长而逐渐增加,作用48h后到最大值,呈现良好量效和时效关系(P<0.05)。其他细胞相关蛋白CYP1B1和CYP3A4没有改变。结论苯并(a)芘可诱导CYP1A1表达增加,可能引起雌激素代谢发生改变,并进而抑制子宫内膜癌细胞的分裂增生。
Objective To investigate the molecular mechanism of benzo (a) pyrene on human estrogen metabolism-related genes in human endometrium. Methods The human endometrial carcinoma cell line was used in vitro to study the effect of benzo (a) pyrene on the cytochrome P450 enzyme activity. RL95-2 cells were treated with 0.016,0.8,0.4,2.0 and 10μmol / L LB for 12h, 2μmol / LBP After treated with RL95-2 cells for 6, 12, 24, 36 and 48h, the cells were harvested and lysed. The protein expression of CYP1A1, CYP1B1 and CYP3A4 in the total cell extracts was detected by Western blot analysis and supersomes expressing CYP450 enzyme as positive control To test the reliability of the method. Results After treated with different doses of BP for 12h, the expression of CYP1A1 protein was significantly increased. After treated with 2μmol / L LB for 2h, the CYP1A1 increased gradually with the prolongation of action time, and reached the maximum after 48h The relationship between dose-effect and aging (P <0.05). Other cell-associated proteins, CYP1B1 and CYP3A4, remained unchanged. Conclusions Benzo (a) pyrene can induce the increase of CYP1A1 expression, which may result in the change of estrogen metabolism and further inhibit the proliferation and proliferation of endometrial cancer cells.