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目的观察放线菌素D(ACTD)对靶细胞和旁观者细胞的光学和超微结构影响,探讨ACTD能否诱导旁观者效应(BE)。方法用不同剂量ACTD(0、0.25、0.5、1.0、2.0、4.0、8.0 mg/L)处理V79靶细胞1 h,在4 mg/LACTD作用4、81、2和24 h取靶细胞去细胞培养液(CM),培养正常细胞24 h后,观察BE;形态学观察用倒置光学显微镜,超微结构用透射电子显微镜观察,存活率用克隆形成实验检测。结果随着ACTD剂量增加,靶细胞数量逐渐减少,而变形细胞数量增多;CM处理旁观者细胞数目减少,脱壁增加;超微结构观察CM处理旁观者细胞出现凋亡征象,光学及超微结构改变均以4 h CM组效应最重,随CM时段延后减轻;细胞存活率在4、8、12、24 h CM组分别为(59.5±3.4)%(、69.2±4.5)%(、88.8±5.2)%和(61.3±6.8)%,均低于对照组,高于4 mg/L ACTD组(50.0±6.5)%(P<0.05)。结论 ACTD可以诱导旁观者细胞凋亡,以4 h CM效应最强。
Objective To observe the optical and ultrastructural effects of actinomycin D (ACTD) on target cells and bystander cells and investigate whether ACTD can induce bystander effect (BE). Methods V79 target cells were treated with different doses of ACTD (0,0.25,0.5,1.0,2.0,4.0,8.0 mg / L) for 1 h. After 4, 81, 2 and 24 h treatment with 4 mg / L ACTD, target cell decellularization Liquid (CM), normal cells were cultured for 24 h, and the morphology of the cells was observed by inverted light microscope. The ultrastructure was observed by transmission electron microscopy. The viability was detected by clone formation assay. Results With the increase of ACTD dose, the number of target cells decreased and the number of deformed cells increased. The number of bystander cells decreased and the number of detached cells increased in CM treated cells. The apoptotic signs were observed in CM treated bystander cells. The optical and ultrastructural changes The changes were all at 4 h CM group was the most significant, with the CM delay postponed; cell survival rate in the 4,8,12,24 h CM group were (59.5 ± 3.4)% (69.2 ± 4.5)% (88.8 ± 5.2% and 61.3 ± 6.8%, respectively, which were lower than those in control group and higher than that in 4 mg / L ACTD group (50.0 ± 6.5)% (P <0.05). Conclusion ACTD can induce bystander cell apoptosis with the strongest CM effect at 4 h.