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目的 探讨抑制热休克蛋白 70 (HSP70 )表达对脑胶质瘤细胞增殖及凋亡的影响。方法 利用细胞培养、电子显微镜及流式细胞仪的方法 ,观察阻断HSP70表达后对细胞生长的影响。结果 体外培养脑胶质瘤细胞株 ,经不同浓度槲皮素作用不同时间后 ,碘化丙碇染色荧光显微镜观察发现细胞核固缩、致密和碎裂现象 ;流式细胞检测发现亚二倍体峰 ,提示出现细胞凋亡。分析细胞周期 ,发现凋亡细胞主要出现在G0 ～G1期 ,随药物浓度的增加而增加 ;观察细胞膜HSP70表达 ,经槲皮素作用后细胞表面HSP70表达显著受到抑制。透射电镜观察凋亡细胞出现核致密、核碎裂及核边聚现象。结论 脑胶质瘤细胞广泛存在HSP70过度表达 ,HSP70可以作为判断肿瘤恶性程度及预后的一个参考指标 ;槲皮素可特异性抑制体外培养的胶质瘤细胞HSP70的表达并诱导凋亡。
Objective To investigate the effect of heat shock protein 70 (HSP70) expression on the proliferation and apoptosis of glioma cells. Methods Cell culture, electron microscopy and flow cytometry were used to observe the effects of blocking HSP70 expression on cell growth. Results In vitro culture of glioma cell lines, different concentrations of quercetin for different periods of time, propidium iodide staining fluorescence microscopy observed nuclear condensation, densification and fragmentation; flow cytometry found subdiploid peaks , suggesting that apoptosis occurs. Analysis of the cell cycle revealed that apoptotic cells mainly appeared in the G0-G1 phase and increased with increasing drug concentration. Observing the expression of HSP70 on the cell membrane, the expression of HSP70 on the cell surface was significantly inhibited by quercetin. Transmission electron microscopy revealed that apoptotic cells exhibited nuclear condensation, nuclear fragmentation, and nuclear condensation. Conclusion HSP70 is widely expressed in glioma cells. HSP70 can be used as a reference to judge the malignancy and prognosis of tumors. Quercetin can specifically inhibit the expression of HSP70 and induce apoptosis in cultured glioma cells.