目的观察β-葡萄糖苷酶激活苦杏仁苷对膀胱癌EJ细胞生长及凋亡的影响,探讨β-葡萄糖苷酶/苦杏仁苷酶解前药系统用于膀胱癌治疗的应用前景。方法用不同浓度的苦杏仁苷与β-葡萄糖苷酶共同作用于膀胱癌EJ细胞株,用MTT法检测细胞的增殖活性,倒置显微镜和透射电镜观察细胞形态学改变,流式细胞仪检测细胞凋亡率与凋亡周期。结果与单纯用苦杏仁苷作用相比,加入β-葡萄糖苷酶后膀胱癌EJ细胞的增殖明显受限,增值抑制率可以达到前者的8.19倍;倒置显微镜和透射电镜观察到肿瘤细胞呈现凋亡的表现,流式细胞仪检测肿瘤细胞凋亡率增加、多数肿瘤细胞阻滞于S期,这些改变随着苦杏仁苷浓度的增加而明显增强。结论β-葡萄糖苷酶激活苦杏仁苷后能够明显抑制EJ细胞的增长,其作用机制可能与将肿瘤细胞阻滞于S期而诱导其凋亡有关;β-葡萄糖苷酶/苦杏仁苷酶解前药系统可能成为治疗膀胱癌的有效策略。
Objective To explore the effects of amygdalin activated by β-glucosidase on the growth and apoptosis of bladder cancer EJ cells,and to discuss β-glucosidase/amygdalin enzyme prodrug system as a potential therapy for bladder cancer.Methods EJ cells were divided into two groups: the amygdalin group,and the β-glucosidase/amygdalin group.After the treatment,the cell growth curve was determined by methyl thiazolyl tetrazolium(MTT).The inverted microscope and transmission electron microscope were used to detect the change of morphology of EJ cells.The apoptosis and cycle of EJ cells were tested by flow cytometry.Results The cell proliferation capability in the β-glucosidase/amygdalin group was significantly inhibited,and the inhibition rate was up to 8.19 times of that in the amygdalin group.The inverted microscope and transmission electron microscope showed a notable apoptosis in the β-glucosidase/amygdalin group.The flow cytometry showed that the β-glucosidase/amygdalin group had a higher apoptosis rate,and more S phase cells.With amygdalin concentration increased,these changes were more obvious.Conclusion Amygdalin activated by β-glucosidase can decrease the proliferation and promote the apoptosis capability of EJ cells,which may explain the reason why more cells were blocked in S phase.β-glucosidase/amygdalin enzyme prodrug system may become a potential therapy for bladder cancer.