Objective To investigate the relationship between Notch signaling pathway and ep-ithelial mesenchymal transition in glioma cells.Methods Immunomagnetic bead was used to isolate brain glioma stem cells from human glioma tissue. The glioma stem cells were cultured in vitro. Immunofluorescence assay was used to identify the cells. RNA interference retroviral vector of Notch-1 gene(pSiRNA-Notch-1) was constructed. The experiment contained three groups: blank control group(non-transfected plasmid), negative control group(transfected with blank plasmid) and interference group(transfected with pSiRNA-Notch-1). The proliferation and differentiation of three groups were observed. The expression of mRNA and protein of Notch-1 and Hes-1 was detected by RT-PCR and Western blot.Results The cell proliferation and cell differentiation absorbance(OD) of interference group after 7d were(1.332±0.614) and(1.203±0.783) respectively, which were significantly lower than that of blank control group and negative control group(P<0.05). The relative expression of mRNA and protein of Notch-1 and Hes-1 in interference group after 3 d proliferation was(0.189±0.021) and (0.301±0.121),(0.422±0.022) and(0.091±0.032), which was significantly lower than that in blank control group and negative control group(P<0.05). The relative expression of mRNA and protein of Notch-1 and Hes-1 in interference group after 3 d differentiation was(0.253±0.071) and(0.192±0.043),(0.178±0.022) and (0.101±0.012), which was significantly lower than that in blank control group and negative control group(P<0.05). Conclusions Notch signaling pathway plays an important role in proliferation and differentiation of glioma cells. Notch-1 and Hes-1 may be involved in the regulation of proliferation and differentiation, which needs further study.

目的 分析NOTCH-1信号通路对人脑胶质瘤干细胞增殖和分化作用.方法 选取医院2015年1月-2017年1月收治的人脑胶质瘤患者50例,从人脑胶质瘤组织标本和细胞株中提出胶质瘤干细胞体外培养,对胶质瘤干细胞增殖和分化作用中NOTCH-1信号通路基因和蛋白表达情况进行分析.结果 高度恶性胶质瘤光密度值(IOD)高于低度恶性胶质瘤(P<0.05);在胶质瘤干细胞增殖过程中,NOTCH-1、CBF-1、HES-1在CD133+中表达较强,干细胞分化时,表达减弱.结论 NOTCH-1信号通路参与了人脑胶质瘤干细胞增殖和分化过程,在人脑胶质瘤中高表达,为疾病治疗提供了重要依据.

目的 探索Notch信号通路在胶质瘤干细胞上皮间充质转化中的意义,以期为临床提供实践价值.方法 选取我院2015年1月~2016年12月间接治的30里人脑胶质瘤手术患者作为本次研究对象,利用免疫磁珠法对人脑胶质瘤干细胞进行提取和分离,并进行体外细胞培养,同时利用免疫组法以及荧光定量PCR对相关指标进行检测,探索Notch信号通路在胶质瘤干细胞上皮间充质转化中机制.结果 经分析对比发现,正常脑组织的NOTCH-1蛋白阳性区域分析发现,其蛋白雷击光密度值为(296.1±28.4),低度恶性胶质瘤患者的NOTCH-1蛋白累积光密度为(5038±119.8),高度恶性胶质瘤患者的NOTCH-1蛋白累积光密度为(12001±137.8).相比于正常脑组织患者而言,高度和低度恶性胶质瘤患者的NOTCH-1蛋白表达水平明显偏高,且高度恶性胶质瘤患者明显高于低度恶性胶质瘤患者NOTCH-1蛋白表达水平,NOTCH-1、CBF-1以及HES-1基因在CD133+中表达水平均明显高于在CD133-胶质瘤细胞,上述差异显著(P<0.05),具有统计学意义.结论 Notch信号通路在胶质瘤干细胞上皮间充质转化中的表达水平高,在人脑胶质瘤干细胞增殖过程中对NOTCH-1、CBF-1以及HES-1等关键基因具有重要的调控作用,对于治疗胶质瘤具有积极的意义.

Objective To observe the expression changes of microRNA181c (miR-181c) in glioblastoma (GBM) and its effects on invasion and migration of tumor cells.Methods real-time PCR was used to detect the miR-181c expression in 41 GBM tissues (GBM group) and 15 normal brain tissues (control group).GBM cell line T98G was divided into four groups:groups A, B, C and D.In the groups A, B and C, the T98G was transfected with miR-181c with overexpression, control miRNA and miR-181c, respectively, but the group D without any treatment.After transfection 48 h, the invasion and migration in each group was investigated.Results Compared with the control group, the miR-181c expression was down-regulated in the GBM group, P<0.05.After transfection 48 h, the invasive cells in the groups B, C and D were more than those of the group A, P<0.05.Moreover, cell relative migration distance in the groups B, C and D was also longer than that of the group A, P<0.05.Conclusion The miR-181c is lowly expressed in GBM tissues, and the low miR-181c expression is closely related with GBM cell invasion and migration.

目的 探讨微小RNA-490-3p(miR-490-3p)对胶质瘤细胞增殖和凋亡的影响,观察miR-490-3p对胶质瘤生物学行为的影响.方法 通过荧光实时定量PCR(qRT-PCR)检测16例胶质瘤样本、胶质瘤旁组织及4种胶质瘤细胞系中miR-490-3p的表达;利用人工合成miR-490-3p mimic瞬时转染脑胶质瘤细胞株,qRT-PCR检测细胞中miR-490-3p的表达水平;采用MTT比色法检测胶质瘤细胞增殖情况;流式细胞术检测胶质瘤细胞凋亡.结果 胶质瘤样本及胶质瘤细胞系中miR-490-3p的表达量较瘤旁与正常胶质细胞系显著降低,miR-490-3p mimic能够显著上调胶质瘤细胞株中miR-490-3p的表达水平,并显著抑制胶质瘤细胞株U251、U87细胞的增殖能力显著增加细胞凋亡数量;结论 miR-490-3p在胶质瘤样本及胶质瘤细胞系中呈现低表达,过表达miR-490-3p有效抑制了胶质瘤细胞株的增殖,增加凋亡,提示miR-490-3p可能成为胶质瘤治疗的新靶点.

Objective To investigate the mechanism of miR -181c inhibiting the invasion of glio‐blastoma cell line T98G .Methods Using RT -PCR and Western Blot to detect the expression of miR -181c and TGF-βrelated genes .The effect of miR -181c and TGF -βon the invasion of glioma cells was measured by Transwell .Using molecular cloning technology to design and clone the target gene 3'-UTR and its mutation region .Using luciferase assay to analyse the combination of miR -181c and target genes .Results mRNA and protein expression of miR -181c and TGF -βpathway associated genes TG‐FBR1 ,TGFBR2 and TGFBRAP1 decreased (P< 0 .05) .miR-181c could weaken the invasion of T98G cells by influencing TGF -βsignal and the 3'-UTR of TGFBR1 ,TGFBR2 and TGFBRAP1 were direct targets of miR-181c .Conclusions miR -181c can inhibit the invasive ability of glioma cell line T 98G by regulating the TGF -βsignaling pathway ,and it can be an important molecule for glioblastoma treatment .