异甘草素对大鼠C6胶质瘤细胞增殖和分化的影响

Effect of Isoliquiritigenin on C6 glioma cell proliferation and differentiation

目的研究异甘草素对大鼠C6胶质瘤细胞增殖和分化的影响。方法采用磺酰罗丹明B(SRB)比色法和克隆形成实验考察异甘草素对C6胶质瘤细胞增殖抑制作用;Giemsa染色观察细胞形态学变化;半定量RT-PCR、Western blot鉴定细胞分化相关基因及蛋白表达水平。结果大鼠C6胶质瘤细胞经异甘草素诱导后,以浓度依赖性抑制细胞增殖(P<0.01);光镜观察到异甘草素诱导前的C6胶质瘤细胞呈两极长梭形或多角形,胞质多,胞突短;而诱导后细胞突起数目不同程度增多,细胞形态变细变长。诱导前克隆形成出现早,数量多,直径大,克隆形成率高;异甘草素处理组克隆形成率降低,与对照组相比差异有统计学意义(P<0.01)。RT-PCR、Western blot结果显示,异甘草素诱导后C6细胞GFAP mRNA和蛋白表达均明显上升(P<0.01)。结论异甘草素能抑制大鼠C6胶质瘤细胞的增殖,并能诱导C6细胞分化。

Aim To investigate the effects of isoliquiritigenin（ ISL） on C6 glioma cell proliferation and differentiation. Methods C6 glioma cells ＇ viability and proliferation were respectively measured by SRB test.Colony formation of C6 glioma cells from different groups was assayed. After culturing the cells from each group,giemsa staining was used to observe cell morphology. RT-PCR was applied to detect mRNA expression of GFAP. Western blot was applied to detect the expression of GFAP. Results ISL effectively inhibited the viability of C6 glioma cells when compared with the control group in a concentration-dependent manner（ P〈0. 01）. The morphological observation under light microscope showed that： in the control group,most of the undifferentiated C6 cells showed long fusiform and polygonal shape. Compared to the control group,the C6 cells treated with ISL revealed alteration in morphology such as astrocytes with smaller smooth,round body and much finer longer,tapering processes. The cloning formation rate detection revealed that： the colonies in the control group semerged earlier and were larger than those experimental ones,the cloning formation rate was higher,while almost no effective cells colony emerged in ISL treated groups（ P〈0. 01）. Western blot and RTPCR analysis showed that GFAP expression in the experimental groups increased（ P〈0. 01）. Conclusion ISL may inhibit the proliferation of C6 glioma cells and induce their differentiation.