芦荟大黄素纳米脂质体介导光动力疗法抑制人口腔癌CAL-27细胞增殖并促进其自噬

Aloe-emodin nanoliposome-mediated photodynamic therapy inhibits proliferation and promotes autophagy of human oral cancer CAL-27 cells

目的:探究芦荟大黄素纳米脂质体(aloe-emodin nanoliposome,AE-L)介导光动力疗法(photodynamic therapy,PDT)对人口腔癌CAL-27细胞的作用效果。方法:逆相蒸发法制备AE-L;MTT法分别检测AE-L(0、2.5、5.0、7.5、10.0、12.5μg/mL)和405 nm激光(光照强度为80 mW/cm2,光照时间为0、10、20、40、80、160 s)对CAL-27细胞存活率的影响;荧光显微镜观察CAL-27细胞摄取AE-L(10μg/mL)情况随时间的变化;MTT法检测AE-L(10μg/mL)介导PDT(光照强度为80 mW/cm2,光照时间0、10、20、40、80、160 s)对CAL-27细胞存活率的影响;用单丹磺酰尸胺(monodansylcadaverine,MDC)荧光染色法和透射电子显微镜观察CAL-27细胞内自噬小体形成情况。结果:CAL-27细胞在6 h摄取AE-L最多,即6 h为PDT光照最佳时间;当AE-L浓度≤10μg/mL时,对CAL-27细胞存活率的影响无明显差异(P>0.05);405 nm激光光照时间不同对CAL-27存活率的影响无明显差异(P>0.05);但AE-L介导PDT能够明显降低CAL-27细胞存活率(P<0.05)且呈光照时间依赖性;AE-L介导PDT组CAL-27细胞产生大量自噬小体,且显著多于对照组。结论:AE-L介导PDT对口腔癌CAL-27细胞增殖具有抑制作用,并且能够促进口腔癌CAL-27细胞发生自噬。

Objective:To investigate the effect of photodynamic therapy(PDT) mediated by aloe-emodin nanoliposome(AE-L) on human oral cancer CAL-27 cells.Methods:AE-L was prepared by inverse evaporation.The effect of AE-L(0,2.5,5.0,7.5,10.0,12.5 μg/mL) or 405 nm laser(power density at 80 mW/cm~2,duration of 0,10,20,40,80,160 s)on the survival rate of CAL-27 cells was analyzed quantitatively by MTT assay.The effects of AE-L and 405 nm laser on the survival rate of CAL-27 cells were detected by MTT method.The uptake of AE-L(10 μg/mL) by CAL-27 cells was observed with fluorescence microscope.The effect of PDT(405 nm laser,power density at 80 mW/cm~2,duration of 0,10,20,40,80,160 s) on the survival rate of CAL-27 cells was analyzed quantitatively by MTT assay.The formation of autophagosomes in CAL-27 cells was detected by using the monodansylcadaverine(MDC) and transmission electron microscopy.Results:The uptake of AE-L by CAL-27 cells reached the maximum at the sixth hour,which was the best time for laser intervention.When the concentrations of AE-L are less than 10 μg/mL,the survival rate of CAL-27 cells was not significantly affected(P>0.05).The effect of 405 nm laser irradiation time on the survival rate of CAL-27 was not significantly different(P>0.05).After treated with the PDT mediated by AE-L,and CAL-27 cells was inhibited in a light-time-dependent manner(P<0.05).Compared with the control group,a large number of autophagosomes were showed in CAL-27 cells of the PDT group.Conclusion:The PDT mediated by AE-L could significantly suppress the proliferation of oral cancer CAL-27 cells and could promote autophagy of oral cancer CAL-27 cells.