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作 者:曾令成[1] 万锋[1] 韩林[1] 叶飞[1] 郭东生[1] 雷霆[1]
机构地区:[1]华中科技大学同济医学院附属同济医院神经外科,武汉430030
出 处:《中华实验外科杂志》2012年第9期1667-1670,共4页Chinese Journal of Experimental Surgery
摘 要:目的探讨CD133mRNA及蛋白表达与胶质瘤细胞未分化状态的相关性。方法对富含AC133表达的胶质瘤干细胞进行了2周诱导分化,实时荧光定量聚合酶链反应(PCR)检测分化前后CD133mRNA表达,Westernblot法检测CD133总蛋白表达,流式细胞学检测细胞表面及细胞内AC133蛋白表达,酶联免疫吸附试验(ELISA)检测培养基上清中AC133蛋白的表达。结果AC133表达随胶质瘤干细胞分化出现显著下调(P〈0.05),CD133mRNA定量表达在NCH421k干细胞中分化前为1.08±0.39,血清环境下分化为1.08±0.51,血清+1μmol/L全反式维甲酸(ATRA)分化下为1.07±0.54,差异无统计学意义(P〉0.05),在NCH441干细胞中分别为2.61±1.72、2.534±1.18、2.18±1.73,差异也无统计学意义(P〉0.05)。CD133总蛋白表达在分化前后无显著改变。结论细胞表面AC133是相对于CD133mRNA及蛋白,能够更敏感地反映胶质瘤细胞未分化状态的指标。Objective To explore the correlation between the expression of CD133 and undifferen- tiated state of glioma cells. Methods Differentiation was induced for 2 weeks in glioma stem cells en- riched with AC133 expression. Real-time quantitative polymerase chain reaction (PCR) was performed to study CD133 mRNA expression. Western blotting was used to detect the CD133 protein expression. By u- sing flow cytometry, extracellular and intracellular AC133 expression was examined. Enzyme linked immu- nosorbent assay (ELISA) was applied to measure the AC133 expression in the culture supernatant. Results The extraceliular AC133 expression in glioma stem cells was significantly down-regulated upon differentia- tion (P〈 0. 05 ). Quantitative CD133 mRNA expression in NCH421k cells before differentiation was 1.08 -+ 0. 39, 1.08 -+ 0. 51 in serum upon differentiation, and 1.07 -+ 0. 54 in serum plus 1 bLmol/L all- trans retinoic acid (P 〉0. 05), and that in NCH441 cells was 2. 61 -+ 1.72, 2. 53 -+ 1.18 and 2. 18 -+ 1.73 respectively (P 〉 0. 05). No significant difference in the CD133 protein expression was observed before and after differentiation. Conclusion Extracelluar AC133 more sensitively mirrors undifferentiated state of glioma cells than CD133 mRNA and protein.
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