本文選題：人腦膠質(zhì)瘤　切入點(diǎn)：CPEB　出處：《現(xiàn)代免疫學(xué)》2017年06期 　論文類型：期刊論文

【摘要】：探討胞質(zhì)多聚腺苷酸化元件結(jié)合蛋白4(cytoplasmic polyadenylation element binding protein 4,CPEB4)調(diào)控NOTCH信號(hào)通路對(duì)腦膠質(zhì)瘤細(xì)胞凋亡的影響。收集人腦膠質(zhì)瘤組織及對(duì)應(yīng)的瘤旁組織,Western blotting檢測(cè)組織中CPEB4蛋白水平。以人腦膠質(zhì)瘤細(xì)胞U87為研究對(duì)象,通過細(xì)胞轉(zhuǎn)染的方法將CPEB4小干擾RNA(CPEB4siRNA)和對(duì)照小干擾RNA(siRNA control)轉(zhuǎn)染至U87細(xì)胞中,同時(shí)設(shè)置對(duì)照組,對(duì)照組細(xì)胞中只加入轉(zhuǎn)染試劑。Western blotting檢測(cè)細(xì)胞中CPEB4水平。流式細(xì)胞儀檢測(cè)細(xì)胞凋亡情況。Western blotting檢測(cè)NOTCH1受體胞內(nèi)段基因NICD1、NOTCH2受體胞內(nèi)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。人腦膠質(zhì)瘤細(xì)胞經(jīng)20μmol/L的NOTCH信號(hào)通路抑制劑S2188作用48h后,檢測(cè)細(xì)胞凋亡及NICD1、NICD2、HES1、Cleaved Caspase-3蛋白水平。人腦膠質(zhì)瘤組織中CPEB4蛋白水平明顯高于瘤旁組織(P0.01)。siRNA control組細(xì)胞中CPEB4、NICD1、NICD2、HES1、Cleaved Caspase-3蛋白水平和細(xì)胞凋亡率與對(duì)照組相比沒有明顯變化(P0.05)。CPEB4siRNA組細(xì)胞中CPEB4水平明顯低于對(duì)照組(P0.01)。CPEB4siRNA組細(xì)胞凋亡率和Cleaved Caspase-3蛋白表達(dá)水平明顯高于對(duì)照組(P0.01)。CPEB4siRNA組細(xì)胞中NICD1、NICD2、HES1蛋白表達(dá)水平明顯低于對(duì)照組(P0.01)。NOTCH信號(hào)通路抑制劑作用后的人腦膠質(zhì)瘤細(xì)胞凋亡情況同CPEB4siRNA組一樣。由此可知CPEB4在人腦膠質(zhì)瘤組織中表達(dá)上調(diào),抑制CPEB4的表達(dá)能夠促進(jìn)人腦膠質(zhì)瘤細(xì)胞凋亡,其作用機(jī)制與NOTCH信號(hào)通路有關(guān)。
[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.
【作者單位】： 鄭州大學(xué)附屬南陽(yáng)醫(yī)院中心醫(yī)院神經(jīng)內(nèi)科;鄭州大學(xué)第二附屬醫(yī)院神經(jīng)內(nèi)科;沈陽(yáng)軍區(qū)總醫(yī)院神經(jīng)內(nèi)科;
【基金】：國(guó)家自然科學(xué)基金(青年科學(xué)基金項(xiàng)目)(81401097)
【分類號(hào)】：R739.41

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