目的：探讨糖原合酶激酶-3β（ GSK-3β）抑制剂TDZD-8（4-苄基-2-甲基-1，2，4-噻二唑烷-3，5-二酮）对大鼠C6胶质细胞瘤细胞增殖的抑制作用及诱导凋亡的机制。方法：采用MTT方法检测TDZD-8不同浓度（10、20、40和80μmol/L）处理大鼠C6胶质细胞瘤细胞6、12、24和48 h后对细胞增殖的影响；流式细胞术检测其对细胞凋亡和细胞周期的作用，同时分析凋亡相关基因 Bax、Bcl-2和P53的表达变化。结果：TDZD-8浓度、时间依赖性的抑制大鼠 C6胶质细胞瘤细胞的增殖（P＜0.05）。流式细胞术显示TDZD-8浓度依赖诱导细胞凋亡，其中在20、40和80μmol/L组有统计学差异（ P＜0.05）。40和80μmol/L TDZD-8处理C6细胞后，G0/G1期细胞数量明显增多，分别升高到83％和90％（ P＜0.05）；而S期和G2/M期细胞数则显著降低（ P＜0.05）。同时，40和80μmol/L TDZD-8使Bax和P53表达显著增加，而使Bcl-2表达降低（ P均＜0.05）。结论：GSK-3β抑制剂TDZD-8可以浓度依赖性抑制C6细胞增殖同时诱导其凋亡，并引起细胞周期阻滞，这可能与其对Bax、P53和Bcl-2的调控相关。%Objective:To investigate the growth-inhibitory and apoptosis-inducing effects and underlying mechanism of TDZD-8,a glycogen synthase kinase-3β( GSK-3β)inhibitor,on rat C6 glioma cells. Methods:MTT method was used to measure the proliferation of rat C6 glioblastoma cells at 6,12,24 and 48h after treated with different concentrations of TDZD-8(10,20 and 80 μmol/L). Flow cytometry was used to detect the cell apoptosis and cell cycles. The expressions of apoptosis-related genes Bax,Bcl-2 and P53 were also analyzed. Results:TDZD-8 was found to inhibit proliferation of rat C6 glioma cells in a concentration-and time-dependent manner( P<0. 05). Flow cytometry showed that TDZD-8 induced cell apoptosis in a concentration-dependent manner,with significant difference among 20,40 and 80μmol/L dose groups( P<0.05). After treatment with 40 and 80 μmol/L TDZD-8,the percentage of G0/G1 phase C6 cells was increased significantly to 83% and 90%, respectively( P<0.05),while the percentage of S and G2/M phase cells was significantly reduced( P<0.05). At the same time,treatment with 40 and 80μmol/L TDZD-8 resulted in significantly increased expression of Bax and P53 and reduced expression of Bcl-2( P<0.05). Conclusion:The GSK-3βinhibitor TDZD-8 may inhibit the C6 cell proliferation in a concentration-dependent manner,and induce apoptosis and cell cycle arrest,which may be related to regulation of Bax,P53 and Bcl-2 expressions.
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