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

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（TYRAP）表达对氢醌诱导人HL．60细胞凋亡的影响，以探讨TYRAP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TYRAP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，TYRAP基因表达量下降。结论体外培养条件下，TYRAP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。

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

OBJECTIVE: To study the effect of TTRAP expression on apoptosis induced by hydroquinone in HL-60 cells in vitro, and explore the relationship between TTRAP 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 TTRAP 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 micromol/L, lasting for 8 h. When it was 250 micromol/L, the necrotic rate increased significantly. The apoptosis induced by hydroquinone was associated with the culture time at the concentration of 200 micromol/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 TTRAP expression in the mRNA level also increased accordingly. When the concentrations of hydroquinone was above 250 micromol/L, necrotic rate increased sharply, and the amount of TTRAP expression decreased. CONCLUSION: Hydroquinone could induce apoptosis of HL-60 cells. The up-regulation of TTRAP expression may promote hydroquinone to induce HL-60 cells to go into apoptosis in vitro with dose-effect and time-effect relationship.