南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (02): 229-.

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替莫唑胺诱导大鼠脑胶质瘤C6细胞caspase非依赖的程序性死亡

刘丽华,张明   

  • 出版日期:2015-02-20 发布日期:2015-02-20

Caspase-independent programmed cell death induced by temozolomide in rat glioma C6
cell line

  • Online:2015-02-20 Published:2015-02-20

摘要: 目的观察替莫唑胺对大鼠脑胶质瘤C6细胞程序性死亡的诱导效应,并探讨可能的分子机制。方法观察不同浓度替莫
唑胺在不同时间点处理大鼠脑胶质瘤大鼠脑胶质瘤C6细胞系,MTT法观察抑制率并筛选出最优的作用时间和作用浓度。应
用400 μg/ml替莫唑胺处理细胞24 h,通过HE染色、Hochest、Western blot、Tunnel、免疫组化观察替莫唑胺对大鼠脑胶质瘤C6
细胞系的程序性死亡的诱导效应。结果MTT法、HE染色、Hochest和Tunnel结果显示替莫唑胺对大鼠脑胶质瘤C6细胞具有
凋亡诱导效应;免疫组化和Western blot结果显示促凋亡蛋白Bax表达水平上调,凋亡抑制蛋白Bcl-2表达的下调。未发现主要
的caspase蛋白的表达变化。结论替莫唑胺对脑胶质瘤C6细胞具有凋亡诱导效应,而这种效应可能是通过caspase非依赖途径
完成的。

Abstract: Objective To investigate programmed cell death induced by temozolomide in rat glioma C6 cell line. Methods Rat
glioma C6 cell line was treated by temozolomide at different concentrations and for different time lengths. MTT assay was
used to evaluate the cell inhibition rate to determine the optimal exposure time and concentration. After the exposure to 400
μg/ml temozolomide for 24 h, the cells were observed for programmed cell death using HE staining, Hochest and Tunnel
assay, Western blotting, and immunohistochemistry. Results MTT, HE staining, and Hochest and Tunnel assay all showed
temozolomide-induced apoptosis in rat glioma C6 cell line. Western blotting and immunohistochemistry revealed
up-regulation of Bax and down-regulation of Bcl-2 expression in the exposed cells, where the expressions of caspases 3, 8, 9,
and 12 remained unchanged. Conclusion Temozolomide induces apoptosis in rat glioma C6 cell line through a caspaseindependent
pathway.