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胶质瘤干细胞是胶质瘤起源和化疗抵抗的重要原因,也是胶质瘤治疗的重要靶点。丙戊酸钠是一种去乙酰化酶抑制剂,具有抗肿瘤活性。本研究主要探讨丙戊酸钠对胶质瘤干细胞生长的影响。我们使用无血清培养方法从A172胶质瘤细胞中,分离培养胶质瘤干细胞,免疫荧光染色发现胶质瘤干细胞表达Nestin和CD133。使用不同浓度(0.5~1.5 mmol/L)丙戊酸钠连续处理胶质瘤干细胞7 d,CCK8检测细胞活力,从第4天起,就可观察到丙戊酸钠处理组细胞OD值显著低于正常组,并且随着丙戊酸钠浓度的增加和处理时间的延长,细胞OD值下降更为明显。检测胶质瘤干细胞克隆形成率,正常组胶质瘤干细胞在干细胞培养基中培养7 d后,克隆形成率为(21±2.03)%,给予0.5 mmol/L丙戊酸钠处理胶质瘤干细胞,克隆形成率明显下降到(13±3.86)%,继续增加丙戊酸钠浓度至1 mmol/L或1.5 mmol/L时,克隆形成率分别非常显著下降至(10±4.71)%和(8±3.66)%。此外,丙戊酸钠也可导致胶质瘤干细胞凋亡,并且细胞凋亡率随着丙戊酸钠浓度的增加而增加。本研究证实丙戊酸钠抑制胶质瘤干细胞增殖和凋亡,并且其抑制效应具有浓度依赖性。丙戊酸钠是一种非常有潜力的治疗胶质瘤的药物,本研究的完成将为它的临床运用提供坚实的理论基础。
Glioma stem cells are an important cause of glioma’s origin and chemotherapy resistance, and also an important target of glioma treatment. Sodium valproate is a sirtuin inhibitor with anti-tumor activity. This study mainly investigated the effect of sodium valproate on glioma stem cell growth. Glioma stem cells were isolated and cultured from A172 glioma cells using serum-free culture method. Nestin and CD133 were expressed in glioma stem cells by immunofluorescence staining. Continuous treatment of glioma stem cells with different concentrations of sodium valproate (0.5 ~ 1.5 mmol / L) for 7 days, CCK8 cell viability detected from the fourth day, the observed valproate sodium group OD value was significantly lower In the normal group, and with the increasing of the sodium valproate concentration and the prolongation of the treatment time, the OD value of the cells decreased more obviously. The colony formation rate of glioma stem cells was detected. After cultured for 7 days in normal group, the colony formation rate was (21 ± 2.03)%, and glioma stem cells were treated with 0.5 mmol / L sodium valproate (10 ± 4.71)% and (10 ± 4.71)%, respectively, when the concentration of sodium valproate was increased to 1 mmol / L or 1.5 mmol / L ± 3.66)%. In addition, sodium valproate also resulted in apoptosis of glioma stem cells, and the rate of apoptosis increased with increasing sodium valproate concentration. This study confirmed that sodium valproate inhibits glioma stem cell proliferation and apoptosis, and its inhibitory effect in a concentration-dependent manner. Sodium valproate is a potential drug for the treatment of glioma. The completion of this study will provide a solid theoretical basis for its clinical application.