李伟, 陈余清, 周继红, 李殿明, 徐凤珍, 夏雪梅. Survivin反义寡核苷酸经线粒体途径诱导肺癌细胞凋亡研究[J]. 蚌埠医学院学报, 2005, 30(6): 482-486.
    引用本文: 李伟, 陈余清, 周继红, 李殿明, 徐凤珍, 夏雪梅. Survivin反义寡核苷酸经线粒体途径诱导肺癌细胞凋亡研究[J]. 蚌埠医学院学报, 2005, 30(6): 482-486.
    LI Wei, CHEN Yu-qing, ZHOU Ji-hong, LI Dian-ming, XU, Feng-zhen. Survivin antisense oligonucleotides induces apoptosis of lung cancer cells through mitochondrial-dependent pathway[J]. Journal of Bengbu Medical College, 2005, 30(6): 482-486.
    Citation: LI Wei, CHEN Yu-qing, ZHOU Ji-hong, LI Dian-ming, XU, Feng-zhen. Survivin antisense oligonucleotides induces apoptosis of lung cancer cells through mitochondrial-dependent pathway[J]. Journal of Bengbu Medical College, 2005, 30(6): 482-486.

    Survivin反义寡核苷酸经线粒体途径诱导肺癌细胞凋亡研究

    Survivin antisense oligonucleotides induces apoptosis of lung cancer cells through mitochondrial-dependent pathway

    • 摘要: 目的: 探讨survivin反义寡核苷酸(antisense oligonucleotides,ASODN)诱导肺癌细胞凋亡的作用及survivin抗凋亡的分子机制。方法: 脂质体介导下,分别以100 nmol/L、300 nmol/L、500 nmol/L survivin ASODN作用于肺癌细胞株NCI-H446后,在不同时间用流式细胞仪(flow cytometry,FCM)检测凋亡;Rh123染色法检测细胞线粒体膜电位(mitochondrial potentialmembrane,△Ψm)变化,EL ISA法检测胞质细胞色素C(cytochrome C,Cyt C)浓度,比色法检测胞质内天冬氨酸特异性半胱氨酸蛋白酶-9(cystenyl aspartate specific proteinase-9,caspase-9)活性变化;加入caspase-9抑制剂Z-LEHD-FMK后FCM检测细胞凋亡。结果: Survivin ASODN 500 nmol/L作用72 h效果最佳,细胞凋亡指数(AI)达48.35%,增殖指数(PI)为24.38%;Survivin ASODN导致细胞△Ψm逐渐下降,并相继引起Cyt C的释放和caspase-9的激活。三者变化具有时间依赖性和差异性;Z-LEHD-FM K显著抑制了survivin ASODN诱导的细胞凋亡。结论: Survivin主要通过调控线粒体凋亡途径发挥抗肺癌细胞凋亡作用;Survivin ASODN能够显著诱导肺癌细胞凋亡,抑制细胞增殖。

       

      Abstract: Objective: To investigate the role of survivin antisense oligonucleotides(ASODN) in inducing the apoptosis of lung cancer cells and its mechanism.Methods: The lung cancer cell lines NCI-H446 were transferred with 100 nmol/L,300 nmol/L and 500 nmol/L survivin ASODN.The apoptosis index(AI)and the proliferation index(PI) were measured by flow cytometry(FCM).The cells were stained with Rh123 to detect the changes in the mitochondrial membrane potential(△Ψm) by FCM.While the concentration of cytoplasmic cytochrome c (Cyt C) was determined by ELISA,the relative activities of caspase-9 were detected by colorimetric assay.The apoptotic rates of lung cancer cells induced by survivin ASODN with or without caspase-9 inhibitor were measured by FCM.Results: When the NCI-H446 cells were treated with concentration of ASODN 500 nmol/L for 72 h,the AI was 48.35%,and the PI 24.38%.Survivin ASODN decreased △Ψm gradually,released Cyt C and activated caspase-9 significantly.The changes of the above indicators had time-dependent and time diversity relationship.In the presence of caspase-9 inhibitor,the apoptotic rates of lung cancer cells induced by survivin ASODN decreased significantly.Conclusions: Survivin inhibits apoptosis primarily via regulation of mitochondrial-dependent pathway.Survivin ASODN can not only induce apoptosis but also inhibit cell proliferation.

       

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