对比硫化砷对两种白血病细胞株凋亡和hTERT-mRNA表达的影响

Comparison of the effect of arsenic sulfide on apoptosis and hTERT-mRNA expression in two leukemia cell lines

目的:对比硫化砷对慢性粒细胞白血病细胞株K562和急性早幼粒细胞白血病细胞株NB4两种细胞的作用是否存在差异及其机制。方法:PCR-ELISA法测定端粒酶活性;半定量RT-PCR检测hTERTmRNA的表达;流式细胞术测量细胞周期、凋亡。结果:0.15～0.6mg/L硫化砷(72h)可在诱导NB4细胞凋亡的同时,下调细胞端粒酶活性和hTERT-mRNA表达,同样作用对于K562细胞需要硫化砷浓度达到0.3～3mg/L。0.3mg/L硫化砷(72h)可引起NB4细胞出现G2/M期细胞比例增高。而K562细胞需要在1.5mg/L硫化砷诱导下出现G2/M期细胞比例增高。结论:端粒酶系统可能是硫化砷诱导NB4和K562细胞凋亡的途径之一;由硫化砷诱导的G2/M期阻滞可能与端粒酶活性的显著下降相关。NB4和K562细胞对硫化砷的敏感性存在明显差异,具体机制有待进一步研究。

AIM： To explore the different effect and mechanism of arsenic sulfide on telemorase activity and hTERT-mRNA expression in CML cell lines-K562 and APL cell lines-NB4. METHODS： Telomerase activity was determined by polymerase chain reaction enzyme-linked immunoassay （PCR-ELISA）. The expression of hTERT-mRNA was analyzed by semi-quantitative RT-PCR. Flow cytometry was used to analyze the cell cycle and apoptosis. RESULTS： 0.1.5 - 0.6 mg/L arsenic sulfide （72 h） can induce apoptosis and inhibit telomerase activity and hTERT-mRNA expression in NB4 cell. The concentration of arsenic sulfide with the same effect on K562 cell was 0.3 - 3 mg/L. 0.3 mg/L arsenic sulfide （72 h） can cause the proportion of the NB4 cell in G2/M phase increased, but for 1〈562 cell, The concentration of arsenic sulfide was 1. 5 mg/L. CONCLUSION： Telomerase system may be one of the pathway for arsenic sulfide inducing apoptosis of NB4 and K562. cell; G2/M phrase arrest may have correlation with decrease of telomerase activity; The sensitivity of NB4 and K562 cell for arsenic sulfide is different, the mechanism of it need to study more.