双酚A暴露对TM3细胞DNA甲基转移酶基因表达影响

Effect of bisphenol A exposure on the expression of DNA methyltransferase gene in TM3 cells

目的探讨双酚A暴露对小鼠睾丸间质细胞株TM3细胞及其DNA甲基转移酶(Dnmt)基因表达的影响。方法将TM3细胞随机分为0、20、50、125、300μmol/L剂量组(0μmol/L剂量组为对照组),予相应浓度的双酚A溶液处理24 h后,采用CCK-8法测定TM3细胞存活率,采用实时荧光定量聚合酶链式反应法检测Dnmt1、Dnmt3a和Dnmt3b mRNA的表达情况。结果随着双酚A染毒剂量的增加,TM3细胞存活率逐渐下降(P<0.01)。随着双酚A染毒剂量的增加,TM3细胞Dnmt1 mRNA相对表达水平下降(P<0.01);20、50、125、300μmol/L剂量组TM3细胞Dnmt3b mRNA相对表达水平均低于对照组(P<0.05)。5组TM3细胞Dnmt3a mRNA相对表达水平比较,差异无统计学意义(P>0.05)。结论双酚A暴露对TM3细胞具有细胞毒性,这种毒性作用可能与Dnmt mRNA表达量发生改变有关。

Objective To investigate the effect of bisphenol A exposure on the expression of DNA methyltransferase(Dnmt)gene in mouse Leydig cell line TM3 cells.Methods TM3 cells were randomly divided into 0,20,50,125 and 300μmol/L dose group(0μmol/L dose group was the control group).Cells were treated with various concentration of bisphenol A solution for 24 hours.The viability of TM3 cells was determined by CCK-8 method,and the mRNA expression of Dnmt1,Dnmt3 a and Dnmt3 b was detected by real-time fluorescence quantitative polymerase chain reaction.ResultsThe viability of TM3 cells decreased with the increasing doses of bisphenol A(P<0.01).The relative mRNA expression of Dnmt1 in TM3 cells decreased with the increase of doses of bisphenol A(P<0.01).The relative mRNA expression of Dnmt3 b in TM3 cells in the 20,50,125,300μmol/L dose group was lower than that in the control group(P<0.05).There was no statistical significant difference in the relative mRNA expression of Dnmt3 a in TM3 cells in these 5 groups(P>0.05).Conclusion Bisphenol exposure is cytotoxic to TM3 cells.This toxic effect may be related to changes in Dnmt mRNA expression.