【摘要】 目的探讨原花青素对髓母细胞瘤Daoy细胞增殖和凋亡的影响及初步机制。方法不同浓度原花青素处理Daoy细胞24、48和72 h,CCK-8检测细胞增殖,Hoechst33258荧光染色法及流式细胞仪分析细胞凋亡,Western blot检测细胞Akt、p-Akt和NF-κB的表达。结果原花青素对肿瘤细胞的增殖抑制效应呈浓度时间依赖性,72 h的半数抑制浓度IC50为0.25 mmol/L。0.2 mmol/L原花青素处理细胞24、48和72 h后,可见较多的凋亡细胞,明显的核固缩;细胞凋亡率分别为21.16%±2.49%、36.02%±0.71%、63.79%±1.47%。细胞总Akt水平无明显变化,但p-Akt和NF-κB蛋白表达水平明显降低。结论原花青素可能通过降低p-Akt和NF-κB的表达,在体外抑制Daoy细胞增殖,并促进其凋亡。更多还原

【Abstract】 Objective To study the effects of proliferation and apoptosis of human medulloblastoma cell line Daoy induced by proanthocyanidins( PC),furthermore to explore the molecular mechanism. Methods Human medulloblastoma cell line Daoy were treated by PC at different concentrations( 0. 025,0. 05,0. 1,0. 2,0. 4 mmol / L) for 24,48 and 72 h,the inhibition rate of tumor cells was determined by CCK-8 assay. Then,the apoptosis of which was evaluated by Hoechst33258 fluorochrome staining and flow cytometry via annexin-V / PI labeling method. The protein expression of Akt,p-Akt,NF-κB was detected by Western blot analysis. Results PC inhibited the proliferation of the Daoy cells in time and dose-dependent manner. After treatment for 72 h,the IC50 of PC was 0. 25 mmol / L. We observed many apoptotic cells that the nuclei were condensed,and flow cytometry found that the apoptotic rates of Daoy cells treated by 0. 2 mmol / L PC for 24,48,72 h were 21. 16% ± 2. 49%,36. 02% ± 0. 71%,63. 79% ±1. 47%,respectively. The results showed that the total Akt protein had not been changed,but the p-Akt and NF-κB protein expression was down-regulated obviously. Conclusions PC inhibits the proliferation of human medulloblastoma cells and induces their apoptosis,and the molecular mechanism is related with down regulation of p-Akt and NF-κB protein expressions in vitro.更多还原