【摘要】 目的:检测胸腺嘧啶激酶基因转染C6胶质瘤细胞后用丙氧鸟苷处理时细胞凋亡的发生。方法:用脂质体介导的基因转染将单纯疱疹病毒胸腺嘧啶激酶基因转入C6胶质瘤细胞中,用G418筛选,建立稳定表达胸腺嘧啶激酶的C6细胞株;加入丙氧鸟苷作用不同时间,用原位末端标记、透射电镜和DNA凝胶电泳法检测。结果:用丙氧鸟苷作用72h发现有大量原位末端标记阳性的细胞,阳性信号呈块状或半月形位于核膜内侧;透射电镜发现肿瘤细胞核内染色质呈块状或帽状位于核膜内侧,非染色质部分呈空泡状,细胞质有的固缩,细胞器基本保持完整;有的则高度肿胀,细胞内亚单位遭到破坏;DNA凝胶电泳发现用丙氧鸟苷作用72h细胞基因组DNA呈梯状带。结论:实验结果提示胸腺嘧啶激酶/丙氧鸟苷系统作用于表达胸腺嘧啶激酶的C6细胞时该细胞主要是以凋亡的方式死亡的。更多还原

【Abstract】 We have examined the mode of cell death in thymidine kinase(tk)/ganciclovior(GCV) killing of C6 glioma cells carrying thymidine kinase gene in vitro by TUNEL, transmission electron microscope and DNA electrophoresis method, tk expression vector carrying neo gene producing resi-sance to G418 was transfected into C6 cells by using Lipofectin and then was selected with G418. tk+-C6-cells were prepared and treated with ganciclovior for different time. TUNEL signals showing DNA fragmentation and chromation condensation (forming either caps or patches narginally to the nuclear membrane) were identified under light microscope in apoptotic cells. U1-trastructual evidence of apoptotic cell death included pyknotic nuclei (condensed chromatin) and apoptotic body formation, however, some of them with cytoplasma swelling and subunit damage. The presence of oligonucleosomal-size fragments on agarose gels was detected. It is concluded that C6 glioma cells carrying tk gene die by a process of apoptosis treated with ganciclovir.更多还原