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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第9期1667-1670,共4页
Chinese Journal of Experimental Surgery
