吉九里香碱诱导K562细胞凋亡的研究

Induction of apoptosis by girinimbine in K562 cell

目的探讨吉九里香碱(girinimbine)通过诱导细胞凋亡而发挥其抑制肿瘤细胞增殖的作用。方法采用MTT法检测吉九里香碱对K562细胞增殖的抑制作用;采用荧光显微镜观察细胞凋亡的形态学变化;库尔特全自动颗粒粒度仪分析药物引起K562细胞体积大小分布的变化;琼脂糖凝胶电泳测定DNA梯形条带及流式细胞术检测细胞凋亡时凋亡峰的变化。结果MTT结果显示不同浓度的吉九里香碱处理K562细胞不同时间均能抑制细胞的增殖,抑制率依赖于吉九里香碱的浓度和作用时间;50μmol/L吉九里香碱处理K562细胞24h时,细胞出现凋亡典型的形态学特征;50μmol/L吉九里香碱分别处理K562细胞6、12、24h及不同浓度吉九里香碱处理K562细胞24h时,能够使细胞产生明显的DNA梯形条带和体积大小变化,呈一定的量效和时效关系,并且细胞凋亡的亚G1峰随作用时间的延长而增加,分别为(15.93±3.79)%(6h)、(32.87±4.89)%(12h)、(41.30±1.91)%(24h)。结论吉九里香碱可能通过诱导K562细胞凋亡而发挥其抗K562细胞增殖的作用。

Objective To investigate the inhibition of girinimbine on cancer cell proliferation by inducing apoptosis. Methods Morphological changes of apoptosis were observed with invert fluorescence microscope; The distribution of the cell volume sizes was assessed by Coulter Multisizer Ⅱ analytical instrument ; DNA Ladder, cell cycle, and the percentage changes of apoptotic cells were examined by DNA agarose gel electrophoresis and flow cytometry （FCM）, respectively. Results MTT Assay showed that girinimbine with different concentrations treated for different times could inhibit K562 cell proliferation. The inhibitory rate was dependent on girinimbine concentrations and treatment times. The K562 cells treated with 50 μmol/L girinimbine for 24 h showed typically morphological changes of apoptotic phase. After treated by 50 μmol/L girinimbine for 6, 12, and 24 h or by different concentrations girinimbine for 24 h, respectively, there were obvious changes of DNA Ladder and the distribution of the cell volume sizes in a time- and dose- dependent manner, and the apoptotic percentage changes of sub-G1 peak cells from （15.93±3.79）% （6 h）, （32.87±4.89）% （12 h）, and （41.30±1.91）% （24 h）, respectively. Conclusion These results suggest that girinimbine maybe show its anticancer activity by inducing apoptosis of K562 cells.