藏红花素对C6胶质瘤细胞生长及Survivin和Livin表达的影响

目的:研究藏红花素对大鼠C6胶质瘤细胞生长及凋亡蛋白抑制因子Survivin和Livin表达的影响。方法:体外培养C6胶质瘤细胞,加入不同浓度的藏红花素培养液,并于不同时间点进行观测,采用四甲基偶氮唑蓝(MTT)比色法绘制细胞生长曲线,观察C6细胞的生长活性;通过相差显微镜和Hoechst荧光染色法观察C6细胞的形态学变化;采用Western blot法检测Survivin和Livin蛋白的表达水平。结果:C6胶质瘤细胞经藏红花素作用后细胞生长受到明显抑制,用含2、4和8 mg/ml藏红花素的培养液作用48h后各组C6细胞的OD值分别为0.732±0.013、0.421±0.010和0.289±0.017,细胞生长抑制率分别为26.8±0.01%、58.0±0.02%和71.1±0.02%,其中4 mg/ml和8 mg/ml藏红花素实验组细胞生长抑制率与阴性对照组均有显著性差异(P均0.05);相差显微镜和Hoechst荧光染色法观察显示实验组C6细胞出现典型的凋亡形态学改变;Western blot检测显示实验组C6细胞Survivin和Livin蛋白表达明显下调。结论:藏红花素能明显抑制C6胶质瘤细胞的体外生长,其抑制作用与诱导C6细胞发生凋亡和下调凋亡蛋白抑制因子Survivin和Livin的表达有关。

Influence of Crocin on Growth and Expression of Survivin and Livin in C6 Glioma Cells

Objective： To study the effect of crocin on growth and expression of inhibitors of apoptosis protein （lAPs） survivin and livin in rat C6 glioma cells. Methods： The C6 glioma cells were cultured in vitro, treated with different concentration of crocin, and ob- served on different time points. The growth activity of C6 cells was investigated using cell growth curve drawn according to methyl thia- zolyl tetrazolium （MTT）-based colorimetric assay. The changes of C6 cells morphology were observed under phase contrast microscope and using Hoechst fluorescent staining. The protein expression level of survivin and livin was analyzed using Western blot. Results： The growth of C6 cells was significantly suppressed in vitro by corcin, at 48 hours after treatment with culture solution including 2, 4 and 8 mg/ml of crocin, the optical density （OD） values of C6 cells were 0.732± 0.013, 0.421±0.010 and 0.289± 0.017 respectively, as well as the growth inhibition rates of C6 cells were 26.8±0.01%, 58.0±0.02% and 71.1± 0.02% respectively, there were significant differences comparing the cell growth inhibition rates of experimental groups with both 4 mg/ml and 8 mg/ml of crocin with that of negative control group （P〈0.05 for both）. The changes of typical apoptotic morphology were presented in C6 cells of experimental groups. In addition, the Western blot analysis showed decreased protein expression of survivin and livin in C6 cells of experimental groups. Conclusion： Corcin can inhibit obviously the growth of C6 cells in vitro, and its inhibition effect is associated with inducing C6 cells apoptosis and down-reg- ulating expression of lAPs surivin and livin in C6 cells.