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目的研究白藜芦醇联合X射线对胶质瘤干细胞辐射抗性的影响。方法采用甲基四唑蓝(MTT)法检测白藜芦醇和X射线对胶质瘤干细胞增殖的抑制作用;光镜下观察胶质瘤干细胞的形态变化;采用Hoechst荧光染色观察凋亡小体;采用单丹磺酞尸胺(MDC)染色和Western blot分析自噬在X射线诱导胶质瘤干细胞死亡中的作用。结果 MTT检测结果显示,与阴性对照组和单独X射线处理组相比,白藜芦醇联合X射线可明显抑制胶质瘤干细胞生长(均P<0.05);在加入自噬抑制剂3-MA后,胶质瘤干细胞抑制率下降(P<0.05)。显微镜下观察联合处理组的细胞形态,细胞不再成球形生长;Hoechst 33258荧光染色结果显示,联合处理组细胞出现凋亡形态;MDC染色结果显示,联合处理组细胞出现大量自噬泡;Western blot分析结果显示,联合处理组细胞的Bcl-2表达减少(P<0.01),LC3Ⅱ/LC3Ⅰ的比值增大(P<0.01)。结论白藜芦醇可提高胶质瘤干细胞的辐射敏感性,白藜芦醇联合X射线处理可诱导胶质瘤干细胞发生凋亡和自噬;自噬对于提高胶质瘤干细胞的辐射敏感性起到重要作用。
Objective To study the effects of resveratrol combined with X-ray on the radiation resistance of glioma stem cells. Methods Methyltetrazolium (MTT) method was used to detect the inhibitory effects of resveratrol and X-ray on the proliferation of glioma stem cells. The morphology of glioma stem cells was observed under light microscope. The apoptotic bodies were observed by Hoechst staining. The role of autophagy in xenograft-induced glioma stem cell death was analyzed by single-stained dichromated phthalocine (MDC) staining and Western blot. Results The results of MTT assay showed that resveratrol combined with X-ray significantly inhibited the growth of glioma stem cells (all P <0.05) compared with the negative control group and X-ray alone group. The addition of autophagy inhibitor 3-MA After glioma stem cell inhibition rate decreased (P <0.05). The morphology of cells in the combined treatment group was observed under a microscope. The cells no longer grew into a spherical shape. Hoechst 33258 fluorescence staining showed that the cells in the combined treatment group showed apoptotic morphology. MDC staining showed that a large number of autophagic vacuoles appeared in the cells treated with the combination of Western blot The results showed that the Bcl-2 expression was decreased (P <0.01) and the ratio of LC3Ⅱ / LC3Ⅰ was increased in the combined treatment group (P <0.01). Conclusion Resveratrol can enhance the radiation sensitivity of glioma stem cells, resveratrol combined with X-ray treatment can induce glioma stem cell apoptosis and autophagy; autophagy to improve glioma stem cell radiation sensitivity To the important role.