Objective: To investigate the effect of kaempferol on proliferation of acute myeloid leukemia (AML) KG1a cells and its mechanism.
Methods: Human AML KG1a cells in logarithmic growth stage were taken and set at 25, 50, 75 and 100 μg/ml kaempferol group, another normal control group (complete medium without drug) and solvent control group (add dimethyl sulfoxide) were also set. After 24 and 48 hours of intervention, the cell proliferation rate was detected by CCK-8 assay. In addition, interleukin-6 (IL-6) combined with kaempferol group (Plus 20 μg/l IL-6 and 75 μg/ml kaempferol) was set up, 48 hours after culture, the cell cycle and apoptosis of KG1a cells were detected by flow cytometry, the mitochondrial membrane potential (MMP) of KG1a cells was detected by MMP detection kit (JC-1 method), and the expression of Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway related proteins in KG1a cells were detected by Western blot.
Results: The cell proliferation rate of 25, 50, 75 and 100 μg/ml kaempferol group decreased significantly (P<0.05), and with the increase of kaempferol dose (r24 h=-0.990, r48 h= -0.999), the cell proliferation rate decreased gradually (P<0.05). The inhibitory effect of 75 μg/ml kaempferol on cell proliferation reached half of effective dose after 48 hours of intervention. Compared with normal control group, the G0/G1 phase cell proportion and apoptosis rate of cells in 25, 50 and 75 μg/ml kaempferol group increased, while the S phase cell proportion, MMP, phosphorylated JAK2 (p-JAK2)/JAK2 and phosphorylated STAT3 (p-STAT3)/STAT3 protein expression decreased in a dose-dependent manner (r=0.998, 0.994, -0.996, -0.981, -0.997, -0.930). Compared with 75 μg/ml kaempferol group, the G0/G1 phase cell proportion and apoptosis rate of cells in IL-6 combined with kaempferol group decreased, while the S phase cell proportion, MMP, p-JAK2/JAK2 and p-STAT3/STAT3 protein expression increased significantly (P<0.05).
Conclusion: Kaempferol can inhibit KG1a cell proliferation and induce KG1a cell apoptosis, its mechanism may be related to the inhibition of JAK2/STAT3 signal pathway.
题目: 山奈素对KG1a细胞增殖的抑制作用及机制研究.
目的: 探讨山奈素对急性髓系白血病（AML）KG1a细胞增殖抑制作用及其相关作用机制。.
方法: 取对数生长期的人AML KG1a细胞，培养液中分别加入山奈素药液0、25、50、75、100 μg/ml，溶剂对照组加入二甲基亚砜，干预24和48 h后，采用CCK-8法检测细胞增殖率；另设白细胞介素-6（IL-6）与山奈素联用组（加20 μg/L的IL-6和75 μg/ml的山奈素），给药培养48 h后，流式细胞术检测KG1a细胞周期及凋亡情况，线粒体膜电位（MMP）检测试剂盒（JC-1法）检测KG1a细胞MMP，蛋白免疫印迹法检测KG1a细胞JAK2/STAT3通路相关蛋白表达。.
结果: 山奈素25、50、75、100 μg/ml组细胞增殖率明显降低（P<0.05），且随着山奈素剂量的增大，细胞增殖率逐渐降低（r24 h= -0.990、r48 h=-0.999）（P<0.05）；75 μg/ml的山奈素干预48 h后对细胞增殖的抑制作用已达到半数有效剂量；与空白对照组相比，山奈素25、50、75 μg/ml组细胞在G0/G1期的细胞比例、凋亡率依次升高，而S期的细胞比例、MMP、磷酸化JAK2（p-JAK2）/JAK2、磷酸化STAT3（p-STAT3）/STAT3蛋白表达依次降低，且呈一定的剂量依赖性（r=0.998、0.994、-0.996、-0.981、-0.997、-0.930）；与山奈素75 μg/ml组相比，IL-6与山奈素联用组细胞在G0/G1期的细胞比例、凋亡率明显降低，S期的细胞比例、MMP、p-JAK2/JAK2、p-STAT3/STAT3蛋白表达明显升高（P<0.05）。.
结论: 山奈素可抑制KG1a细胞增殖，诱导KG1a细胞凋亡，其作用机制可能与抑制JAK2/STAT3信号通路有关。.
Keywords: Janus kinase 2/signal transducer and activator of transcription 3; acute myeloid leukemia; apoptosis; kaempferol; proliferation.