赖氨匹林对C6胶质瘤细胞的增殖抑制和促凋亡作用

Anti-proliferation and apoptosis effects of aspisol on C6 glioma cells

目的研究赖氨匹林（aspis01）对体外培养的C6胶质瘤细胞的增殖抑制以及促凋亡作用。方法采用四氮唑（MTr）比色法测定aspisol对C6细胞的增殖抑制作用；荧光显微镜、透射电镜进行细胞凋亡形态观察；RT—PCR、Western—blot进行凋亡相关基因Box、Bcl-xl mRNA和蛋白表达检测。结果Aspisol对C6细胞的增殖具有抑制作用；aspisol（0．1ug／mL）作用24h使细胞发生典型的凋亡形态改变；BoxmRNA表达水平随aspisol作用时间的延长而升高，Bcl-xl mRNA表达水平随aspisol作用时间的延长而下降，6h、12h、24h3个时间点表达水平差异有统计学意义（P〈0．05）：Bax蛋白表达水平随aspisol作用时间的延长而升高，Bcl—xl蛋白表达水平随aspisol作用时间的延长而下降，6h与12h、24h3个时间点表达水平差异有统计学意义（P〈0．05）。结论Aspisol对C6细胞具有增殖抑制以及促凋亡作用，推测aspisol通过Bax／Bcl—xl途径诱导C6细胞凋亡。

Objective To investigate the anti-proliferation and apoptosis effects oflysine aspirin （aspisol） on in vitro cultured C6 glioma cells. Methods MTT colorimetric assay was employed to determine the anti-proliferation effect of aspisol on C6 cells. Fluorescence microscopy and transmission electron microscopy were used to observe the morphological changes of apoptosis. The mRNA and protein expressions of apoptosis-related genes Box and Bcl-xl were detected by RT-PCR and Western blotting. Results Aspisol inhibited the proliferation of C6 cells. Typical apoptotic morphological changes were showed when the cells were treated with 0.1 ug/mL aspisol for 24 h. The mRNA expression level of Box was elevated and that of Bcl-xl was declined following the increased processing time; the mRNA expression levels of Bax and Bcl-xl among 6, 12 and 24 h of treatment with 0.1 ug/mL aspisol were significantly different （P〈0.05）. The protein expression level of Box was elevated following the increased processing time and significant difference was noted among 6, 12 and 24 h of treatment with aspisol （P〈0.05）. The protein expression level of Bcl-xl was declined following the increased processing time and significant difference was noted among 6, 12 and 24 h of treatment with aspisol （P〈 0.05）. Conclusion Aspisol can inhibit the proliferation of C6 cells and may induce the cell apoptosis through Box/Bcl-xl channel.