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目的研究三氧化二砷(As2O3)对人脑胶质瘤干祖细胞增殖和周期的影响及其相关分子表达。方法采用四甲基偶氮唑盐(MTT)法测定As2O3对人脑胶质瘤干祖细胞抑制率,流式细胞术检测As2O3对该细胞凋亡和周期的影响,透射电镜观察细胞形态学变化,Western blot和RT-PCR检测Fas、FasL和p53变化。结果 As2O3可明显抑制人脑胶质瘤干祖细胞的增殖,且呈一定的时间剂量依赖关系(P<0.05);此外,人脑胶质瘤干祖细胞周期G1期细胞的比例增加,S期和G2/M期细胞的比例减少;通过形态学观察发现,随着As2O3剂量的增加,镜下凋亡细胞比例逐渐增加,凋亡小体的数量增加;Western blot检测发现,FasL表达上调,p53下调(P<0.05)。结论 As2O3对人脑胶质瘤干祖细胞具有抑制增殖和诱导凋亡作用,其机制可能与上调FasL和下调p53表达相关。
Objective To study the effects of arsenic trioxide (As2O3) on the proliferation and the cycle of human glioma stem and progenitor cells and their related molecular expressions. Methods The inhibitory rate of As2O3 on human glioma stem and progenitor cells was determined by MTT assay. The effect of As2O3 on the apoptosis and cell cycle was assayed by flow cytometry. The morphological changes of cells were observed by transmission electron microscopy The changes of Fas, FasL and p53 were detected by Western blot and RT-PCR. Results As2O3 could significantly inhibit the proliferation of human glioma stem and progenitor cells in a dose-dependent manner (P <0.05). In addition, the proportion of cells in G1 phase of human glioma stem-progenitor cells increased, And the proportion of G2 / M phase cells decreased. The morphological observation showed that with the increase of As2O3 dosage, the proportion of apoptotic cells increased gradually and the number of apoptotic bodies increased. Western blot showed that FasL expression was up-regulated and p53 Down (P <0.05). Conclusion As2O3 can inhibit proliferation and induce apoptosis of human glioma stem and progenitor cells. The mechanism may be related to the up-regulation of FasL and down-regulation of p53 expression.