肿瘤坏死因子受体相关因子及蛋白表达对氢醌诱导HL-60细胞凋亡的影响

Effect of TTRAP expression on apoptosis induced by hydroquinone in HL-60 cells in vitro

目的研究体外培养条件下肿瘤坏死因子受体相关因子及蛋白(TTRAP)表达对氢醌诱导人HL-60细胞凋亡的影响,以探讨TTRAP表达与氢醌诱导HL-60细胞凋亡的关系。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;反转录聚合酶链反应(RT-PCR)方法检测TTRAP在mRNA水平的表达量,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、12h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,差异有统计学意义(P<0.01),氢醌诱导细胞凋亡的最佳浓度为200μmol/L,而当氢醌浓度为250μmol/L时,细胞坏死率明显增高,浓度为20μmol/L氢醌诱导的细胞凋亡,随着作用时间的延长,凋亡率明显增高,作用8h达高峰,而后细胞凋亡率下降,坏死率增加。作用8h时,随着氢醌浓度的增加,细胞凋亡率明显增加,TTRAP基因在mRNA表达水平也相应明显增加;当浓度增加到250μmol/L时,细胞坏死率增加,TTRAP基因表达量下降。结论体外培养条件下,TTRAP的表达上调可能对氢醌诱导HL-60细胞凋亡起促进作用,并存在剂量-效应与时间-效应关系。

Objective To study the effect of TYRAP expression on apoptosis induced by hydroquinone in HL-60 cells in vitro, and explore the relationship between TYRAP expression and the apoptosis. Methods Apoptotic and necrotic rate was examined by flow cytometer with Anti-AnnexinV/FITC Plus PI staining. The mRNA expression of TYRAP was detected by RT-PCR. The differences in different treated groups were compared. Results After different concentrations of hydroquinone to the cells for 0, 4, 8, 12 h culture, were added, the cell apoptotic rate in different concentrations of hydroquinone groups was significantly higher than that in blank control groups. The optimal concentration of hydroquinone was 200 μmol/L, lasting for 8 h. When it was 250μmol/L, the necrotic rate increased significantly. The apoptosis induced by hydroquinone was associated with the culture time at the concentration of 200 μmol/L, and the peak apoptotic time was 8 h. Then the apoptotic rate decreased and necrotic rate increased. Furthermore, with the concentrations of hydroquinone increased and time lasted for 8 h, the apoptotic rate of cells increased ,the amount of TYRAP expres- sion in the mRNA level also increased accordingly. When the concentrations of hydroquinone was above 250 μmol/L, necrotic rate increased sharply, and the amount of TYRAP expression decreased. Conclusion Hydroquinone could induce apoptosis of HL-60 cells. The up-regulation of TYRAP expression may promote hydro- quinone to induce HL-60 cells to go into apoptosis in vitro with dose-effect and time-effect relationship.