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为研究单胺氧化酶抑制剂(TCP)对体外培养的人脑胶质瘤U251细胞的诱导分化作用,以不同浓度TCP单独或与100nmol/L组蛋白去乙酰化酶抑制剂(TSA)联合处理U251细胞。采用倒置显微镜观察细胞形态学变化;噻唑蓝(MTT)比色法检测细胞增殖变化;流式细胞仪检测细胞周期变化;Hoechst 33258染色显示细胞凋亡;Real-time PCR和Western印迹法检测分化相关基因mRNA和蛋白表达水平的改变。结果表明:TCP可诱导细胞突起增多且变细长,抑制细胞增殖,阻滞细胞周期于G1期,抑制全能性转录因子Oct4和Sox2的表达,上调分化标志基因胶质纤维酸性蛋白(GFAP)的表达,TCP联合TSA也诱导了GFAP的高表达。这些结果提示:TCP可诱导胶质瘤U251细胞分化,TSA对TCP诱导细胞分化有协同作用。
U251 cells were treated with different concentrations of TCP alone or in combination with 100 nmol / L histone deacetylase inhibitor (TSA) in order to study the induction of differentiation of human glioma U251 cells by monoamine oxidase inhibitor (TCP). The morphological changes of the cells were observed by inverted microscope. The cell proliferation was detected by MTT colorimetric assay. The cell cycle was detected by flow cytometry. The apoptosis was detected by Hoechst 33258 staining. The expression of apoptosis was detected by Real-time PCR and Western blot Gene mRNA and protein expression levels change. The results showed that TCP could induce more prominence and slender cells, inhibit cell proliferation, arrest the cell cycle in G1 phase, inhibit the expression of pluripotency transcription factors Oct4 and Sox2, and upregulate the differentiation marker glial fibrillary acidic protein (GFAP) Expression, TCP combined with TSA also induced high GFAP expression. These results suggest that TCP can induce glioma U251 cells to differentiate and that TSA has a synergistic effect on TCP-induced cell differentiation.