胞质多聚腺苷酸化元件结合蛋白4调控NOTCH信号通路对脑胶质瘤细胞凋亡的影响

发布时间：2018-03-03 05:38

本文选题：人脑胶质瘤　切入点：CPEB　出处：《现代免疫学》2017年06期 　论文类型：期刊论文

【摘要】：探讨胞质多聚腺苷酸化元件结合蛋白4(cytoplasmic polyadenylation element binding protein 4,CPEB4)调控NOTCH信号通路对脑胶质瘤细胞凋亡的影响。收集人脑胶质瘤组织及对应的瘤旁组织,Western blotting检测组织中CPEB4蛋白水平。以人脑胶质瘤细胞U87为研究对象,通过细胞转染的方法将CPEB4小干扰RNA(CPEB4siRNA)和对照小干扰RNA(siRNA control)转染至U87细胞中,同时设置对照组,对照组细胞中只加入转染试剂。Western blotting检测细胞中CPEB4水平。流式细胞仪检测细胞凋亡情况。Western blotting检测NOTCH1受体胞内段基因NICD1、NOTCH2受体胞内段基因NICD2、NOTCH通路下游靶基因HES1、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved cysteinyl aspartate specific proteinase 3,Cleaved Caspase-3)蛋白水平。人脑胶质瘤细胞经20μmol/L的NOTCH信号通路抑制剂S2188作用48h后,检测细胞凋亡及NICD1、NICD2、HES1、Cleaved Caspase-3蛋白水平。人脑胶质瘤组织中CPEB4蛋白水平明显高于瘤旁组织(P0.01)。siRNA control组细胞中CPEB4、NICD1、NICD2、HES1、Cleaved Caspase-3蛋白水平和细胞凋亡率与对照组相比没有明显变化(P0.05)。CPEB4siRNA组细胞中CPEB4水平明显低于对照组(P0.01)。CPEB4siRNA组细胞凋亡率和Cleaved Caspase-3蛋白表达水平明显高于对照组(P0.01)。CPEB4siRNA组细胞中NICD1、NICD2、HES1蛋白表达水平明显低于对照组(P0.01)。NOTCH信号通路抑制剂作用后的人脑胶质瘤细胞凋亡情况同CPEB4siRNA组一样。由此可知CPEB4在人脑胶质瘤组织中表达上调,抑制CPEB4的表达能够促进人脑胶质瘤细胞凋亡,其作用机制与NOTCH信号通路有关。
[Abstract]:To investigate the effect of cytosolic polyadenosylated polyadenylation element binding protein 4 (CPEB4) on the apoptosis of glioma cells regulated by NOTCH signal pathway. Western blotting was collected for the detection of CPEB4 protein in human glioma and its adjacent tissues. Taking human glioma cell line U87 as the object of study, CPEB4 small interfering RNAs (CPEB4 siRNAs) and control small interfering RNA(siRNA (RNA(siRNA control) were transfected into U87 cells by cell transfection. In the control group, only transfection reagent was added. Western blotting was added to detect the level of CPEB4. Flow cytometry was used to detect the apoptosis of cells. Western blotting was used to detect the intracellular segment of NOTCH1 receptor gene NICD1 and NOTCH2 receptor gene. Cysteine aspartate protein hydrolase 3cleaved cysteinyl aspartate specific proteinase 3Caspase-3. Human glioma cells were treated with 20 渭 mol/L NOTCH signal pathway inhibitor S2188 for 48 h. The level of CPEB4 protein in human glioma tissue was significantly higher than that in the adjacent tissue (P0.01n.siRNA control group). There was no significant change in the protein level and apoptosis rate of CPEB4NICD1 + NICD1 + HES1Cleaved Caspase-3 compared with the control group. CP4siRNA group had no significant change in the level of CPEB4 protein. CP4siRNA group had no significant change compared with the control group. The level of CPEB4 was significantly lower than that of the control group (P0.01C. CPEB4siRNA) and the expression of Cleaved Caspase-3 protein was significantly higher than that of the control group (P0.01n.CPEB4siRNA group). The expression level of NICD1pNICD2HES1 protein was significantly lower than that of the control group treated with P0.01n.NOTCH signal pathway inhibitor. The expression of CPEB4 was up-regulated in human glioma tissues. Inhibiting the expression of CPEB4 can promote the apoptosis of human glioma cells, and its mechanism is related to the NOTCH signaling pathway.
【作者单位】：郑州大学附属南阳医院中心医院神经内科;郑州大学第二附属医院神经内科;沈阳军区总医院神经内科;
【基金】：国家自然科学基金(青年科学基金项目)(81401097)
【分类号】：R739.41

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