苯丁酸钠体外诱导脑胶质瘤细胞分化的实验研究

Sodium phenylbutyrate(SPB) induces differentiation of the C6 glioma cells in vitro:an experimental study

目的探讨苯丁酸钠(SPB)对脑胶质瘤C6细胞的增殖抑制、诱导凋亡作用。方法C6细胞分别经1、2、4 mmol/L SPB处理后,应用光镜观察细胞形态学变化;电镜观察细胞超微结构变化;MTT法检测细胞增殖情况;流式细胞仪检测细胞周期;软琼脂克隆形成率检测法测定克隆形成率;免疫组织化学染色检测GFAP表达。结果C6胶质瘤细胞的光镜下、电镜下的结构改变显示细胞分化趋于成熟;MTT显示胶质瘤细胞增殖受到抑制;各浓度SPB处理组细胞处于G0/G1期细胞数增加(P<0.01),而处于S期细胞数减少(P<0.01),4 mmol/L SPB处理组可见凋亡峰;软琼脂克隆形成率检测示克隆形成率降低(P<0.01);GFAP表达增多。结论苯丁酸钠可明显抑制C6细胞增殖,诱导细胞分化,甚至诱发细胞凋亡。

Objective To investigate the effects of sodium phenylbutyrate （SPB） on proliferation inhibition and apoptosis induction of the brain glioma cells. Methods The C6 glioma cells were first induced by 1, 2, 4 mmol/L SPB, then the morphological changes of the C6 cells were observed by a hght microscope; the ultra-structure of the cells were observed by an electron microscope; cell proliferation was determined by MTT assay; cell cycle was analyzed by flow cytometry; cell colony formation rate was measured by the soft agar clone forming test; and expression of GFAP was determined by immanohistochemistry staining. Results The morphology and ultra-structure of the C6 cells were differentiated towards normal ceils; cell proliferation was inhibited by MTT assay; cells that stayed at the G0/G1 phage were increased （ P 〈 0.01）, while those that stayed at the S phage were decreased （ P 〈 0.01）, and cell apoptosis peaked in the 4 mmol/L SPB group as shown by flow cytometry; cell cloning efficiency was degraded （ P 〈 0.01） by the soft agar clone forming test; and expression of GFAP was increased. Conclusions SPB can apparently inhibit the proliferation of C6 cells and can induce cell differentiation and even apoptosis of C6 cells.