蓬子菜总黄酮诱导NB4细胞凋亡作用及机制

朱玉良; 马英丽; 李瑞; 张紫阳; 董均明; 马玲

doi:10.11847/zgggws2017-33-02-10

蓬子菜总黄酮诱导NB4细胞凋亡作用及机制

Pro-apoptotic effect of FGVL on NB4 cells and its mechanism

摘要

摘要: 目的观察蓬子菜总黄酮（FGVL）诱导急性早幼粒细胞白血病细胞株（NB4）细胞凋亡作用，并探讨其作用机制。方法 FGVL（50、100、200 μg/mL）作用于体外培养的NB4细胞，Hoechst染色观察细胞形态学变化；流式细胞术检测细胞周期，Annexin V-FITC/PI双标记法检测细胞凋亡状况；RT-PCR检测NB4凋亡相关基因Mel-18、Sall4、Bmi-1 mRNA表达。结果细胞形态学观察显示，NB4细胞经FGVL处理48 h后，与对照组比较，FGVL各剂量组NB4细胞凋亡比例均明显升高（P<0.05），呈现细胞核固缩、核碎裂及凋亡小体等典型的细胞凋亡特征；与对照组（2.3%）比较，50、100、200 μg/mL FGVL组NB4细胞凋亡率（分别为9.3%、13%、20.4%）明显升高（P<0.05）；细胞周期检测结果显示，与对照组比较，50、100、200 μg/mL FGVL组NB4细胞G0/G1期细胞比例（分别为43.92%、37.26%、29.15%）减少，S期细胞比例（54.16%、62.23%、70.46%）增多；与对照组比较，100、200 μg/mL FGVL组NB4细胞Mel-18 mRNA表达水平升高，Sall4、Bmi-1 mRNA表达水平明显降低。结论 FGVL可阻滞NB4细胞于S期，抑制细胞增殖，通过调节Mel-18、Sall4、Bmi-1等凋亡相关基因表达诱导细胞凋亡。

Abstract: Objective To observe pro-apoptotic effect of flavone from Galium verum L (FGVL) on acute promyelocytic leukemia cells (NB4) and to explore the mechanism of the effect.Methods NB4 cells in culture medium in vitro were treated with different concentrations of FGVL (50,100,and 200 μg/mL).Changes in cell morphological were observed with Hoechst staining and the changes in cell cycle and apoptosis rate were analyzed with flow cytometry.The expression levels of Mel-18,Sall4,and Bmi-1mRNA were assessed with reverse transcriptase PCR(RT-PCR).Results Morphological observations revealed that the percentages of apoptotic cells were significantly enhanced in NB4 cells treated with FGVL for 48 hours,with the percentages of 9.3%,13%,and 20.4% (P<0.05 for all) for the dosages of 50,100,and 200 μg/mL groups,compared with that of in the control group (2.3%);typical apoptotic morphological changes (karyopyknosis,karyorrhexis,and apoptosis bodies) were observed in the NB4 cells.Cell cycle analysis indicated that compared with the control group,the cell ratios in G0/G1 phase (43.92%,37.26%,ans 29.15%) decreased significantly,while the ratios in S phase (54.16%,62.23%,and 70.46%) increased obviously for the NB4 cells treated with 50,100,and 200 μg/mL FGVL.Compared with the control group,the expression level of Mel-18 mRNA increased and Sall4 and Bmi-1mRNA decreased obviously for the NB4 cells treated with 100 and 200 μg/mL FGVL.Conclusion FGVL could inhibit cell proliferation,induce cell apoptosis,and arrest cells in S phase in NB4 cells;the mechanism of the effects is probably through regulating the expressions of Mel-18,Sall4,and Bmi-1 gene.

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