凋亡;_维生素K2;_HL60细胞株;

凋亡相关基因survivin、bcl2和bax的表达在维生素K2诱导HL60细胞凋亡中的作用

http://www.100md.com 《现代医学》 2006年第5期

凋亡;,维生素K2;,HL60细胞株;,survivin;,bcl2;,bax,,]凋亡;,维生素K2;,HL60细胞株;,survivin;,bcl2;,bax,凋亡相关基因survi

     [摘要]目的探讨维生素K2(VK2)诱导急性早幼粒细胞白血病(AML)细胞株HL60细胞凋亡的作用机制。方法采用透射电镜技术观察VK2处理后细胞结构变化；流式细胞术AnnexinVFITC/PI分析细胞凋亡；PI染色分析细胞周期变化； RTPCR技术检测凋亡相关基因survivin、bcl2和bax的表达。结果 40μmol·L-1VK2作用HL60细胞72h后，电镜下可见典型细胞凋亡的形态改变；流式细胞术结果显示随着VK2浓度的增加和作用时间的延长细胞凋亡率逐渐增高并呈明显的浓度、时间依赖，与对照组相比差异有显著性 (P<005)；同时S期细胞逐渐减少，G0/G1期细胞逐渐增多(P<005)，细胞被阻滞在G0/G1期；且随着VK2浓度的增加抗凋亡基因survivin、bcl2表达明显下调(P<005)，而促凋亡基因bax表达无显著差异(P>0.05)。结论 VK2能诱导HL60细胞发生凋亡，抗凋亡基因survivin和bcl2mRNA下调可能是VK2诱导HL60细胞发生凋亡的机制之一。

    [关键词]凋亡; 维生素K2; HL60细胞株; survivin; bcl2; bax

    Expressions of apoptosisrelated gene survivin,bcl2 and

    bax in the VK2induced apoptosis of HL60 cells

    SHAO Zeye1, CHEN Baoan2, XIA Guohua1, ZHU Huaigang1,ZHANG Xianwei1,

    HE Zeng1, SHANG Yu1,CHEN Xiaoling1(1 Department of Clinical Laboratory, School of Clinical Medicine，Southeast University，Nanjing 210009，China；

    2 Department of Hematology, Zhongda Hospital,Southeast University, Nanjing 210009, China)

    Abstract: Objective To study the effects and possible mechanisms of the apoptosis of HL60 cells induced by vitamin K2(VK2)Methods The transmission electron microscope was used to observe the changes of morphologic features of HL60 cellsThe possible mechanisms of the apoptosis of HL60 cells induced by VK2 were investigated with the flow cytometry(FCM). Cell cycle changes were analysed by PI staining. Expressions of survivin, bcl2, bax were detected by reverse transcriptase polymerase chain reaction(RTPCR)Results Apoptosis peak on FCM and positive AnnexinVFITC on cell membrane showed that VK2 induced apoptosis of HL60 cells and in a doseandtimedependent manner, and the apoptosis rate was higher than that in contrd group. Cells were arrested in G0/G1 phase, cells in G0/G1 phase were more than those in S phase(P<0.05). The expression of bcl2 and survivin were significantly downregulated(P<0.05), and the expression of bax had no change(P>0.05)Conclusion VK2 could induce apoptosis of HL60 cells, downregulation of the apoptosis related genes bcl2 and survivin might play an important role in this process ......

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