自噬在白藜芦醇调节胶质瘤干细胞辐射敏感性中的作用  

Autophagy involved in Resveratrol increased radiosensitivity in glioma stem cells

作　　者：龙林梅[1] 张庆青[1] 杨能[1] 季文君[1] 宋云珍[1] 赵江虎[1] 梁中琴[1] 

机构地区：[1]苏州大学医学部药理学系,江苏苏州215123

出　　处：《苏州大学学报（医学版）》2012年第2期149-152,169,共5页Suzhou University Journal of Medical Science

基　　金：国家自然科学基金(30873052;81072656);江苏省自然科学基金(BK2010226)资助项目;江苏高校优势学科建设工程资助项目

摘　　要：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 investigate the effect of Resveratrol combined with X-ray on radiosensitivity in glioma stem cells. Methods The proliferation inhibition of glioma stem cells induced by X-rays and Resveratrol was assessed with MTT assay. The activation of proapoptotic effect was characterized by Ho- echst 33258 stain. MDC stain and Western blot analysis were used to analyze the autophagy mechanism in X-rays-induced death of glioma stem cells. Results MTT assay indicated that X-rays and Resveratrol de- creased the viability of glioma stem cells （P 〈 0.05 ） ; we found the proliferative inhibition of glioma stem cells was declined when we used 3-MA to inhibit autophagy（P 〈0.05）. When the cells were treated by the Resveratrol and x-rays, their spherical shape were changed. Apoptosis was induced in glioma stem cells by combined X-rays and Resveratrol as detected by Hoechst 33258 staining. In addition, autophagy was induced in glioma stem cells in the combined treatment group as detected by MDC staining. Western blotting showed that Bcl-2 expression was decreased in the combined treatment group （P 〈 0.01 ） , and the LC3-II expression was increased in the combined treatment group （ P 〈 0.01 ）. Conclusion Resvera- trol can increased the radiation sensitivity of glioma stem cells, the apoptosis and autophagy was induced in the glioma stem cells in the combined treatment X-rays and Resveratrol. Our results suggest that autoph- agy plays an essential role in the regulation of radiosensitization of glioma stem cells.

关 键 词：胶质瘤干细胞 X射线 白藜芦醇 凋亡 自噬 

分 类 号：R811.5[医药卫生—放射医学]