长春新碱诱导的自噬性凋亡与细胞内游离Ca~(2+)浓度的相关性研究

目的　研究长春新碱 (VCR)诱导的L 0 2细胞自噬性凋亡时细胞内游离钙离子浓度 (Ca2 +]i)的变化 ,以及自噬特异性抑制剂 3 methyladenine(3MA)对此自噬性凋亡和 Ca2 +]i的影响。方法　应用已建立的VCR诱导L 0 2细胞自噬性凋亡模型 ,使用电镜、流式细胞术检测细胞凋亡 ;用Fluo 3/AM荧光探针经流式细胞仪测定L 0 2细胞平均 Ca2 +]i。结果　电镜及流式细胞术检测证实VCR诱导L 0 2细胞发生了自噬性凋亡 ,此凋亡过程中 Ca2 +]i明显升高 ,以凋亡早期更为明显 :3MA可显著抑制VCR所致的 Ca2 +]i升高并能降低凋亡细胞比例。结论　在体外条件下VCR可以诱导L 0 2细胞自噬性凋亡 ,其发生可能与VCR导致 Ca2 +]i升高有关 ;3MA可能通过抑制 Ca2 +]i升高而抑制自噬及VCR所致的自噬性凋亡。

Association study on autophagic apoptosis induced by vincristine in cell line L-02 and the intracellular free Ca~(2+)

Objective To study the change of intracellular free calcium levels(Ca 2+ ]i)in the process of autophagic apoptosis induced by vincristine(VCR) in cell line L 02 and to research the influence of 3 methyladenine(3MA), a special inhibitor of autophagy,on the autophagic apoptosis and the intracellular free calcium levels.Methods Human liver cell line L 02 was used as in vitro models.Apoptosis induced by VCR in L 02 cells was investigated by electron microscopy and flow cytometry.Ca 2+ ]i was determined by applying staining of Fluo 3 fluorescein,which fluorescent intensities were measured by flow cytometry.Results Vincristine can not only induced L 02 cells autophagic apoptosis,but also elevate Ca 2+ ]i,Ca 2+ ]in the process of autophagic apoptosis of L 02 cells was significantly higher than that in control groups,especially at the beginning stage of the apoptosis.In group treated with VCR plus 3MA,mean Ca 2+ ]i and the apoptotic cells ratios was significantly lower than that treated with VCR alone.Conclusion The increase of Ca 2+ ]i in VCR treated L 02 cells may be related to autophagic apoptosis induced by VCR.3MA may reduce VCR induced apoptosis by inhibiting increase of Ca 2+ ]i and by inhibiting autophagy in L 02 cells.