论文部分内容阅读
目的探讨丙戊酸钠诱导人脑胶质瘤细胞分化时作用的靶细胞和靶分子。方法丙戊酸钠治疗位于裸小鼠皮下的可移植性人脑胶质瘤,半定量 RT-PCR 检测移植瘤中 HDAC1和 Tob 的表达;CD133免疫磁珠分离胶质瘤组织中的 CD133~+细胞,在分别含血清或不含血清加生长因子培养的条件下,加丙戊酸钠作用21 d,流式细胞仪检测和共聚焦显微镜观察分化标志物表达。结果丙戊酸钠能明显抑制位于裸小鼠皮下移植瘤的增殖(P<0.05),使 HDAC1表达下降(P<0.01),Tob 表达升高(P<0.05)。流式细胞仪检测丙戊酸钠作用21 d 的细胞,在含血清条件下,GFAP 或β-tubulin Ⅲ阳性细胞数显著增加(P<0.01),而无血清加生长因子条件下两标志物表达均无显著差异(P>0.05);共聚焦显微镜观察显示上述的 GFAP~+或β-tubulin Ⅲ~+细胞共表达 Nestin。结论丙戊酸钠逆转人脑胶质瘤细胞分化抑制的靶分子有 HDAC1和 Tob,靶细胞为处于分化过程中的脑肿瘤干细胞。
Objective To investigate the effect of sodium valproate on the differentiation of human glioma cells target cells and target molecules. Methods Sodium valproate was used to treat transplanted human glioma subcutaneously in nude mice. The expression of HDAC1 and Tob in the transplanted tumor was detected by semi-quantitative RT-PCR. CD133 + The cells were treated with sodium valproate for 21 days under the condition of serum-free or serum-free plus growth factor respectively. The expression of differentiation markers was observed by flow cytometry and confocal microscopy. Results Sodium valproate significantly inhibited the proliferation of subcutaneous xenografts in nude mice (P <0.05), decreased the expression of HDAC1 (P <0.01) and increased the expression of Tob (P <0.05). Flow cytometry was used to detect the effect of sodium valproate on cells for 21 days. The number of GFAP or β-tubulin Ⅲ positive cells increased significantly (P <0.01) in serum-containing condition, while the expression of two markers in serum-free plus growth factor (P> 0.05). Confocal microscopy showed that Nestin was co-expressed by GFAP ~ + or β-tubulin Ⅲ ~ + cells. Conclusion Sodium valproate reverses the differentiation of human glioma cells with HDAC1 and Tob and the target cells are brain tumor stem cells in the process of differentiation.