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目的探讨自噬在光敏剂5-氨基乙酰丙酸(aminolaevulinic acid,ALA)介导的光动力治疗(photo-dynamic therapy,PDT)C6胶质瘤细胞死亡中作用机制。方法ALA-PDT处理后不同时间,用自噬特异染料MDC和免疫荧光法观察自噬水平变化,Western blotting检测自噬相关蛋白cathepsin B的表达水平。结果MDC染色和免疫荧光观察可见ALA-PDT后C6胶质瘤细胞内有大量含点状荧光颗粒的自噬泡和自噬体标志物LC3绿色荧光蛋白,Western-blot示自噬相关蛋白cathepsin B表达水平明显升高,其自噬水平随ALA-PDT后时间而逐渐增强。结论ALA-PDT可诱导C6胶质瘤细胞发生自噬,其诱导胶质瘤细胞发生自噬的机制可能与溶酶体酶cathepsin B大量释放有关。
Objective To investigate the mechanism of autophagy in the apoptosis of C6 glioma cells induced by photosensitizer 5-aminolevulinic acid (ALA), a photodynamic therapy (PDT). Methods At different time points after ALA-PDT treatment, autophagy-specific MDC and immunofluorescence were used to observe the changes of autophagy. Western blotting was used to detect the expression of autophagy-related protein cathepsin B. Results The results of MDC staining and immunofluorescence showed that there were a large number of autophagic vacuoles and autophagosome marker LC3 green fluorescent protein in C6 glioma cells after ALA-PDT. Western-blot showed that the autophagy-related protein cathepsin B The level of autophagy increased with time after ALA-PDT. Conclusions ALA-PDT can induce autophagy in C6 glioma cells and its mechanism of inducing autophagy in glioma cells may be related to the massive release of lysosomal enzyme cathepsin B.