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目的研究氢醌对体外培养条件下MOLT4细胞的毒性作用及细胞色素P450CYP4F3基因表达的意义。方法流式细胞术ANNEXINV-FITC加碘化丙啶(PI)双染定量检测细胞凋亡率和坏死率的变化;RT-PCR方法检测CYP4F3在mRNA表达水平,比较不同处理组之间的差异。结果加入不同浓度氢醌培养0、4、8、12、16 h后,流式细胞术检测结果发现,不同浓度氢醌作用后,细胞凋亡率明显高于空白对照组,差异有显著性(P<0.01),氢醌诱导细胞凋亡的最佳时间是8h,而随着作用时间延长,氢醌浓度为200μmol/L时,细胞坏死率明显增高;另外发现氢醌浓度为120μmol/L时,细胞凋亡率随着时间的延长逐步增加,8 h达到高峰,随后开始下降;CYP4F3基因在mRNA表达水平与细胞凋亡率一致。结论体外培养条件下,CYP4F3基因的表达上调可能对氢醌诱导MOLT4细胞凋亡起促进作用,并存在一定量-效与时-效关系。
Objective To study the toxic effects of hydroquinone on MOLT4 cells in vitro and the significance of cytochrome P450CYP4F3 gene expression. Methods Flow cytometry was used to detect apoptosis rate and necrosis rate by ANNEXINV-FITC and PI staining. The mRNA expression level of CYP4F3 was detected by RT-PCR, and the differences among different treatment groups were compared. Results After adding different concentrations of hydroquinone for 0, 4, 8, 12 and 16 h, the results of flow cytometry showed that the apoptotic rate of cells treated with different concentrations of hydroquinone was significantly higher than that of the blank control group P <0.01). The optimal time of hydroquinone-induced apoptosis was 8h. With the prolongation of time, the cell necrosis rate was significantly increased when the concentration of hydroquinone was 200μmol / L. In addition, when the concentration of hydroquinone was 120μmol / L , The rate of apoptosis increased gradually with time, peaked at 8 h, and then began to decline; CYP4F3 gene mRNA expression levels consistent with the rate of apoptosis. Conclusion Upregulation of CYP4F3 gene may promote the apoptosis of MOLT4 cells induced by hydroquinone, and there is a certain amount - effect and time - effect relationship.