STAT3信号转导通路抑制剂诱导胶质瘤干细胞产生自噬的研究

Induction of Autophagy in GSC by a STAT3 Pathway Inhibitor

背景与目的:胶质瘤干细胞(glioma stem cell,GSC)在胶质瘤发展及治疗抗拒中发挥重要作用。我们以往的研究表明,新型STAT3信号转导通路抑制剂(STAT3 inhibitor,STI)WP1193能够诱导GSC产生细胞周期阻滞及凋亡。本研究旨在探讨STI是否能在体外诱导GSC产生自噬现象。方法:从手术切除的胶质母细胞瘤中分离及培养GSC。使用STI处理GSC。利用细胞计数法检测STI对GSC增殖的影响。使用Western blot检测自噬相关蛋白LC3的表达情况。吖啶橙染色后,利用荧光显微镜及流式细胞技术检测酸性自噬小体。使用透射电镜检测GSC中自噬小体。结果:STI剂量依赖性的抑制GSC的增殖。STI处理后,GSC中出现LC3表达的切换。STI处理后,GSC中出现自噬小体,且出现自噬小体细胞的比例增加。结论:STI能够在GSC中诱导自噬现象的产生。自噬在STI治疗中的意义及调节机制需要进一步的研究。

BACKGROUND OBJECTIVE： Glioma stem cells（GSC） are considered as the culprit of the progression and treatment resistance in gliomas.Our previous study demonstrated that novel STAT3 inhibitor（STI） WP1193 induces cell cycle arrest and apoptosis in GSCs.In this study,we aimed to explore whether STI can induce autophagy in GSCs.METHODS： GSCs were isolated and maintained from freshly surgical glioma samples.The effect of STI on GSC proliferation was investigated with cell counting.The expression of autophagy-associated protein LC3 was detected with Western blot.Autophagosomes were detected with acridine orange staining and electron microscopy.RESULTS： STI inhibited the proliferation of GSCs in a dose-dependent manner.LC3-I to LC3-II conversion was found in GSCs after STI treatment.STI induced the accumulation of autophagosomes in GSC.CONCLUSIONS： STI treatment induces autophagy in GSCs.The mechanism of autophagy induced by STI in GSCs needs further investigation.