异甘草素联合替莫唑胺对SHG44人脑胶质瘤干细胞增殖的影响

Effects of Isoliquiritigenin in combination with Temozolomide on cell proliferation of SHG44 human brain glioma stem cells

目的 探讨异甘草素联合替莫唑胺对SHG44人脑胶质瘤干细胞增殖的影响。方法 采用含无血清达尔伯克氏改良伊格尔氏培养基（DMEM/F12）（含表皮生长因子（EGF）、成纤维细胞生长因子（FGF）、B27）的悬浮细胞培养板培养、纯化胶质瘤干细胞，并通过CD133联合Nestin标记进行免疫荧光鉴定；实验分为对照组／[含二甲基亚砜（DMSO）／]、异甘草素（5~160 μmol/L）组、替莫唑胺（12.5～200 μmol/L）组、异甘草素＋替莫唑胺组（160 μmol/L＋12.5~200 μmol/L），通过细胞增殖毒性检测（CCK-8）法、流式细胞术、免疫荧光染色法分别检测细胞抑制率、细胞周期及干细胞表面相关分化蛋白表达情况。结果异甘草素随着浓度增加细胞抑制作用越强，且160 μmol/L时抑制率达32%，替莫唑胺随着浓度增加抑制作用越强，且200 μmol/L时抑制率达40%；异甘草素联合替莫唑胺组随着浓度增加抑制作用越强，且160 μmol/L＋200 μmol/L时抑制率达72%；随着异甘草素浓度增加G0/G1期细胞比例增加，细胞周期阻滞于G0/G1期；随着异甘草素浓度增加干细胞分化越多，且分化细胞的死亡越多。结论异甘草素能诱导SHG44人脑胶质瘤干细胞向星形胶质细胞和神经元细胞分化，且能抑制增殖，同时增强替莫唑胺化疗敏感性。

Objective The effect of Isoliquiritigenin in combination with Temozolomide on cell proliferation of SHG44 human glioma stem cells are investigated.MethodsGlioma stem cells were cultured in serumfree medium supplemented with epidermal growth factor （EGF）, fibroblast growth factor （FGF） and B27, and the suspension cell culture plate was used. CD133 and Nestin were detected using immunofluorescence analysis. The experiment included control group （dimethyl sulfoxide, DMSO）, different concentration of isoliquiritin group （5~160 μmol/L）, temozolomide group （12.5~200 μmol/L）, isoliquiritin＋temozolomide group （160 μmol/L＋12.5~200 μmol/L）. The cell inhibition rates, the cell cycle and stem cell surface differentiated protein expression were detected by cell counting Kit8 （CCK8） method, flow cytometry and immunofluorescence.ResultsThe cell inhibition rate was increased with the isoliquiritin concentration increasing, and the inhibition rate was 32% when the concentration reached 160 μmol/L. The cell inhibition rate was increased with the temozolomide concentration increasing, and the inhibition rate was 40% when the concentration reached 200 μmol/L. The cell inhibition rate was increased with the increase of concentration of isoliquiritin in combination with temozolomide, and the inhibition rate reached 72% when the concentration reached 160＋200 μmol/L. The cell cycle G0/G1 phase was increased with the increase of the concentration of isoliquiritin, and the cell cycle was arrested in G0/G1 phase. With the increase of the isoliquiritin concentration, the more stem cell differentiation, the more cell death.ConclusionIsoliquiritigenin can induce SHG44 glioma stem cells into astrocytes and neuron cell differentiation, and can inhibit the proliferation of glioma stem cells, and increase the chemotherapy sensitivity to temozolomide.