高级搜索

S-Trityl-L-Cysteine诱导HL-60细胞有丝分裂阻滞和凋亡

S-Trityl-L-Cysteine Induces Mitotic Arrest and Apoptosis of HL-60 Cell

  • 摘要: 目的 研究 S-三苯甲基-L-半胱氨酸(S-Trityl-L-Cysteine,STLC)对急性白血病 HL-60细胞有丝分裂阻滞和凋亡的影响,初步探讨药物作用下HL-60细胞有丝分裂阻滞和凋亡间的因果关系。 方法 将HL-60细胞分成不加药对照组和不同剂量 (1、2.5、5、10、50、100 μmol/L) STLC加药组,药物作用一定时间后,台盼蓝染色观察HL-60细胞活性变化, MTT法检测其对HL-60细胞的抑制效应, 免疫荧光染色观察细胞及其核的特征,流式细胞术分析细胞周期和亚二倍体峰的变化,Annexin-v/PI双染检测药物处理前后正常骨髓细胞凋亡比率。 结果 MTT和台盼蓝染色实验显示STLC抑制HL-60细胞生长并致其死亡;免疫荧光染色见核破裂及凋亡小体和细胞体积增大等典型有丝分裂灾变细胞凋亡现象;细胞周期和凋亡分析表明 STLC 致HL-60细胞凋亡比率与药物浓度和作用时间呈正相关,致G2/M期阻滞比率24 h和48 h随药物浓度增加而升高,而72 h无明显变化,进一步研究发现STLC处理早期即可致G2/M期有丝分裂阻滞和凋亡的同时发生,撤除药物作用后有丝分裂阻滞呈可逆性恢复,STLC作用下正常骨髓细胞凋亡比率无明显变化。 结论 STLC诱导HL-60细胞阻滞在G2/M期并致其凋亡,显示较强的抗有丝分裂和抗肿瘤效果,药物处理早期有丝分裂阻滞和凋亡同时发生提示尚有该药物作用新机制的存在。

     

    Abstract: Objective To investigate the effect of S-Trityl-L-Cysteine on the mitosis arrest and apoptosis of acute leukemia HL-60 cell,and approach the causality of mitosis arrest and apoptosis in HL-60 cell under the drug treatment. Methods HL-60 cell was exposed to various concentrations of STLC (1,2.5,5,10,50,100 μmol/L) and compared with the control group.The inhibitory effects was detected by MTT assay, and Trypan blue staining assay was used to examine the number of cell activity. Cell cycle arrest and sub-G1 was detected by flow cytometry assay and character of cell and its nuclear were observed by immunofluorescence staining, Annexin-v/PI assay was used to annlyse the apoptosis rates of normal bone marrow cells treated by STLC. Results MTT and Trypn blue staining assay revealed that STLC inhibited HL-60 cell growth and induced its death.The typical phenomenon of mitotic catastrophe was observed by immunofluorescence staining.STLC induced apoptosis on HL-60 in dose and time dependent and with the augment of STLC, the rate of G2/M arrest increased in 24h and 48h, but no significant change in 72h.Further study revealed G2/M arrest and apoptosis were cencurrent in early stage of drug treament. The mitotic arrest was reversibly after the drug removed. The apoptosis rates of normal bone marrow cells was slightly increase under the drug treatment. Conclusion S-Trityl-L-Cysteine induces G2/M arrest and apoptosis in HL-60 cell and manifest more potential effects on antimitotsis and antitumour. Simultaneous of G2/M arrest and apoptosis suggests that there is an possible new mechanism in the early stage of drug treatment.

     

/

返回文章
返回